#METABOLOMICS WORKBENCH Jung_DNLI_20240919_115027 DATATRACK_ID:5207 STUDY_ID:ST003501 ANALYSIS_ID:AN005749 PROJECT_ID:PR002147 VERSION 1 CREATED_ON September 26, 2024, 4:43 pm #PROJECT PR:PROJECT_TITLE TREM2 expression level is critical for microglial state, metabolic capacity and PR:PROJECT_TITLE efficacy of TREM2 agonism PR:PROJECT_TYPE Preclinical Mouse study PR:PROJECT_SUMMARY Triggering receptor expressed on myeloid cells 2 (TREM2) is a central regulator PR:PROJECT_SUMMARY of microglial activity and sequence variants are major risk factors for late PR:PROJECT_SUMMARY onset Alzheimer’s disease (LOAD). To better understand the molecular and PR:PROJECT_SUMMARY functional changes associated with TREM2 signalling, we generated a TREM2 PR:PROJECT_SUMMARY reporter mouse model and observed a gradual upregulation of reporter expression PR:PROJECT_SUMMARY with increasing plaque proximity. Isolated microglia were sorted based on PR:PROJECT_SUMMARY reporter expression and their transcriptomic profiles acquired in both wildtype PR:PROJECT_SUMMARY and APP transgenic animals, allowing us to disentangle TREM2 versus PR:PROJECT_SUMMARY pathology-specific effects. Bulk RNA sequencing highlighted TREM2 PR:PROJECT_SUMMARY level-dependent changes in major immunometabolic pathways, with enrichment of PR:PROJECT_SUMMARY genes in oxidative phosphorylation and cholesterol metabolism in microglia with PR:PROJECT_SUMMARY increased TREM2 expression. To confirm these findings, we next analysed uptake PR:PROJECT_SUMMARY of fluorodeoxyglucose (FDG) and examined metabolomic and lipidomic profiles. PR:PROJECT_SUMMARY Again, independent of Aβ pathology, TREM2 expression correlated with uptake of PR:PROJECT_SUMMARY FDG as well as increased cellular redox, energetics, and cholesterol PR:PROJECT_SUMMARY homeostasis. Finally, we performed chronic treatment with a brain penetrant PR:PROJECT_SUMMARY TREM2 agonist and identified a window of TREM2 expression where microglia are PR:PROJECT_SUMMARY most responsive. Thus, our data provide novel insights into TREM2-mediated PR:PROJECT_SUMMARY regulation of microglial metabolic function and informs current efforts to bring PR:PROJECT_SUMMARY TREM2 agonists into clinical application. PR:INSTITUTE Denali Therapeutics PR:DEPARTMENT Development Sciences PR:LAST_NAME Suh PR:FIRST_NAME Jung PR:ADDRESS 161 Oyster Point Blvd, South San Francisco, California, 94080, USA PR:EMAIL suh@dnli.com PR:PHONE +1 6507973837 PR:FUNDING_SOURCE Denali Therapeutics PR:PUBLICATIONS https://www.biorxiv.org/content/10.1101/2024.07.18.604115v1 #STUDY ST:STUDY_TITLE TREM2 expression level is critical for microglial state, metabolic capacity and ST:STUDY_TITLE efficacy of TREM2 agonism ST:STUDY_SUMMARY Triggering receptor expressed on myeloid cells 2 (TREM2) is a central regulator ST:STUDY_SUMMARY of microglial activity and sequence variants are major risk factors for late ST:STUDY_SUMMARY onset Alzheimer’s disease (LOAD). To better understand the molecular and ST:STUDY_SUMMARY functional changes associated with TREM2 signalling, we generated a TREM2 ST:STUDY_SUMMARY reporter mouse model and observed a gradual upregulation of reporter expression ST:STUDY_SUMMARY with increasing plaque proximity. Isolated microglia were sorted based on ST:STUDY_SUMMARY reporter expression and their transcriptomic profiles acquired in both wildtype ST:STUDY_SUMMARY and APP transgenic animals, allowing us to disentangle TREM2 versus ST:STUDY_SUMMARY pathology-specific effects. Bulk RNA sequencing highlighted TREM2 ST:STUDY_SUMMARY level-dependent changes in major immunometabolic pathways, with enrichment of ST:STUDY_SUMMARY genes in oxidative phosphorylation and cholesterol metabolism in microglia with ST:STUDY_SUMMARY increased TREM2 expression. To confirm these findings, we next analysed uptake ST:STUDY_SUMMARY of fluorodeoxyglucose (FDG) and examined metabolomic and lipidomic profiles. ST:STUDY_SUMMARY Again, independent of Aβ pathology, TREM2 expression correlated with uptake of ST:STUDY_SUMMARY FDG as well as increased cellular redox, energetics, and cholesterol ST:STUDY_SUMMARY homeostasis. Finally, we performed chronic treatment with a brain penetrant ST:STUDY_SUMMARY TREM2 agonist and identified a window of TREM2 expression where microglia are ST:STUDY_SUMMARY most responsive. Thus, our data provide novel insights into TREM2-mediated ST:STUDY_SUMMARY regulation of microglial metabolic function and informs current efforts to bring ST:STUDY_SUMMARY TREM2 agonists into clinical application. ST:INSTITUTE Denali Therapeutics ST:LAST_NAME Suh ST:FIRST_NAME Jung ST:ADDRESS 161 Oyster Point Blvd, South San Francisco, California, 94080, USA ST:EMAIL suh@dnli.com ST:PHONE +1 6507973837 ST:NUM_GROUPS 5 ST:TOTAL_SUBJECTS 26 ST:NUM_MALES 26 ST:STUDY_COMMENTS Release as soon as it is possible ST:PUBLICATIONS https://www.biorxiv.org/content/10.1101/2024.07.18.604115v1 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:AGE_OR_AGE_RANGE 15 mo SU:GENDER Male #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS Pool_1 HSA-000019615 genotype:NA | condition:NA | group:NA | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_002; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_001; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_002; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_002 SUBJECT_SAMPLE_FACTORS 2_59_mid_WT HSA-000019571 genotype:WT | condition:mid | group:mid_healthy | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_003; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_009; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_003; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_003 SUBJECT_SAMPLE_FACTORS 12_63_mid_WT HSA-000019581 genotype:WT | condition:mid | group:mid_healthy | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_004; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_027; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_004; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_004 SUBJECT_SAMPLE_FACTORS 24_78_low_APP HSA-000019593 genotype:APP | condition:low | group:low_disease | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_005; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_017; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_005; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_005 SUBJECT_SAMPLE_FACTORS 10_62_mid_WT HSA-000019579 genotype:WT | condition:mid | group:mid_healthy | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_006; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_016; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_006; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_006 SUBJECT_SAMPLE_FACTORS 15_69_low_WT HSA-000019584 genotype:WT | condition:low | group:low_healthy | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_007; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_022; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_007; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_007 SUBJECT_SAMPLE_FACTORS 25_78_mid_APP HSA-000019594 genotype:APP | condition:mid | group:mid_disease | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_008; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_012; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_008; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_008 SUBJECT_SAMPLE_FACTORS 18_74_mid_APP HSA-000019587 genotype:APP | condition:mid | group:mid_disease | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_009; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_003; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_009; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_009 SUBJECT_SAMPLE_FACTORS 17_74_low_APP HSA-000019586 genotype:APP | condition:low | group:low_disease | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_010; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_004; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_010; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_010 SUBJECT_SAMPLE_FACTORS 8_61_high_APP HSA-000019577 genotype:APP | condition:high | group:high_disease | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_011; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_011; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_011; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_011 SUBJECT_SAMPLE_FACTORS 21_76_mid_WT HSA-000019590 genotype:WT | condition:mid | group:mid_healthy | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_012; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_020; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_012; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_012 SUBJECT_SAMPLE_FACTORS 14_65_mid_WT HSA-000019583 genotype:WT | condition:mid | group:mid_healthy | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_013; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_028; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_013; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_013 SUBJECT_SAMPLE_FACTORS 7_61_mid_APP HSA-000019576 genotype:APP | condition:mid | group:mid_disease | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_014; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_023; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_014; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_014 SUBJECT_SAMPLE_FACTORS 1_59_low_WT HSA-000019570 genotype:WT | condition:low | group:low_healthy | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_015; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_026; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_015; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_015 SUBJECT_SAMPLE_FACTORS 5_60_high_APP HSA-000019574 genotype:APP | condition:high | group:high_disease | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_016; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_007; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_016; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_016 SUBJECT_SAMPLE_FACTORS 22_77_low_WT HSA-000019591 genotype:WT | condition:low | group:low_healthy | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_017; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_010; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_017; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_017 SUBJECT_SAMPLE_FACTORS Pool_2 HSA-000019616 genotype:NA | condition:NA | group:NA | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_018; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_014; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_018; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_018 SUBJECT_SAMPLE_FACTORS 23_77_mid_WT HSA-000019592 genotype:WT | condition:mid | group:mid_healthy | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_019; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_015; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_019; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_019 SUBJECT_SAMPLE_FACTORS 11_63_low_WT HSA-000019580 genotype:WT | condition:low | group:low_healthy | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_020; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_019; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_020; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_020 SUBJECT_SAMPLE_FACTORS 3_60_low_APP HSA-000019572 genotype:APP | condition:low | group:low_disease | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_021; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_008; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_021; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_021 SUBJECT_SAMPLE_FACTORS 26_78_high_APP HSA-000019595 genotype:APP | condition:high | group:high_disease | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_022; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_025; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_022; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_022 SUBJECT_SAMPLE_FACTORS 19_74_high_APP HSA-000019588 genotype:APP | condition:high | group:high_disease | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_023; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_013; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_023; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_023 SUBJECT_SAMPLE_FACTORS 9_62_low_WT HSA-000019578 genotype:WT | condition:low | group:low_healthy | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_024; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_002; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_024; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_024 SUBJECT_SAMPLE_FACTORS 6_61_low_APP HSA-000019575 genotype:APP | condition:low | group:low_disease | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_025; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_021; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_025; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_025 SUBJECT_SAMPLE_FACTORS 20_76_low_WT HSA-000019589 genotype:WT | condition:low | group:low_healthy | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_026; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_024; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_026; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_026 SUBJECT_SAMPLE_FACTORS 4_60_mid_APP HSA-000019573 genotype:APP | condition:mid | group:mid_disease | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_027; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_006; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_027; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_027 SUBJECT_SAMPLE_FACTORS 13_65_low_WT HSA-000019582 genotype:WT | condition:low | group:low_healthy | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_028; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_018; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_028; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_028 SUBJECT_SAMPLE_FACTORS 16_69_mid_WT HSA-000019585 genotype:WT | condition:mid | group:mid_healthy | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_029; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_005; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_029; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_029 SUBJECT_SAMPLE_FACTORS Pool_3 HSA-000019617 genotype:NA | condition:NA | group:NA | Sample source:microglia RAW_FILE_NAME(Raw Source Name.LP)=20220209_BL_HEX-000330_Lipid_Pos_030; RAW_FILE_NAME(Raw Source Name.TG)=20220324_BL_HEX-000330_TGs_Pos_029; RAW_FILE_NAME(Raw Source Name.MP)=20220209_BL_HEX-000330_Met_Pos_030; RAW_FILE_NAME(Raw Source Name.MN)=20220211_BL_HEX-000330_Met_Neg_030 #COLLECTION CO:COLLECTION_SUMMARY Microglia isolation was performed using the Magnetic-activated cell sorting CO:COLLECTION_SUMMARY (MACS) system (Miltenyi Biotec). No CD11b enrichment step was performed as the CO:COLLECTION_SUMMARY intrinsic mKate2 reporter is only expressed in TREM2 expressing (microglia) CO:COLLECTION_SUMMARY cells. Hence, fluorescence-activated cell sorting was performed based on the CO:COLLECTION_SUMMARY mKate2 fluorescence signal and sorted into low, mid and high expressing CO:COLLECTION_SUMMARY subpopulations, as a proxy for the TREM2 expression of individual cells. CO:SAMPLE_TYPE microglia CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY No treatment #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Pellets from the sorted subpopulations were reconstituted on ice in 9:1 SP:SAMPLEPREP_SUMMARY MeOH:water including internal standards, vortexed for 1 minute, and spun down SP:SAMPLEPREP_SUMMARY for 5 minutes at 10,000 g. Supernatant was transferred to glass vial for SP:SAMPLEPREP_SUMMARY analysis by LC-MS. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY 5 µL of sample was injected on an Imtakt Intrada Organic Acid 3 µm, 2 x 150 mm CH:CHROMATOGRAPHY_SUMMARY (Imtakt USA, Portland, OR USA); using a flow rate of 0.2 mL/min at 60°C. For CH:CHROMATOGRAPHY_SUMMARY negative ionization mode, Mobile phase A consisted of acetonitrile/water/formic CH:CHROMATOGRAPHY_SUMMARY acid = 10/90/0.1%. Mobile phase B consisted of acetonitrile/ 100mM ammonium CH:CHROMATOGRAPHY_SUMMARY formate= 10/90%. The gradient was programmed as follows: 0.0–1.0 min at 0% B; CH:CHROMATOGRAPHY_SUMMARY 1.0–7.0 min to 100% B; 7.1 at 0% B; and 7.1-10 min at 0% B. CH:CHROMATOGRAPHY_TYPE Ion exchange CH:INSTRUMENT_NAME Agilent 1290 Infinity II CH:COLUMN_NAME Imtakt Intrada Organic Acid (150 x 2mm, 3um) ) CH:SOLVENT_A 10% acetonitrile/90% water; 0.1% formic acid CH:SOLVENT_B 10% acetonitrile/90% water; 100mM ammonium formate CH:FLOW_GRADIENT 0.0–1.0 min at 0% B; 1.0–7.0 min to 100% B; 7.1 at 0% B; and 7.1-10 min at CH:FLOW_GRADIENT 0% B. CH:FLOW_RATE 0.20 mL/min CH:COLUMN_TEMPERATURE 60 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex 6500+ Qtrap MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS curtain gas at 40 V; collision gas was set at medium; ion spray voltage at -4500 MS:MS_COMMENTS V; temperature at 600°C; ion source Gas 1 at 50 psi; ion source Gas 2 at 60 MS:MS_COMMENTS psi; entrance potential at -10 V; and collision cell exit potential at -15.0 MS:MS_COMMENTS V. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS log2(area) MS_METABOLITE_DATA_START Samples HSA-000019615 HSA-000019571 HSA-000019581 HSA-000019593 HSA-000019579 HSA-000019584 HSA-000019594 HSA-000019587 HSA-000019586 HSA-000019577 HSA-000019590 HSA-000019583 HSA-000019576 HSA-000019570 HSA-000019574 HSA-000019591 HSA-000019616 HSA-000019592 HSA-000019580 HSA-000019572 HSA-000019595 HSA-000019588 HSA-000019578 HSA-000019575 HSA-000019589 HSA-000019573 HSA-000019582 HSA-000019585 HSA-000019617 Factors genotype:NA | condition:NA | group:NA | Sample source:microglia genotype:WT | condition:mid | group:mid_healthy | Sample source:microglia genotype:WT | condition:mid | group:mid_healthy | Sample source:microglia genotype:APP | condition:low | group:low_disease | Sample source:microglia genotype:WT | condition:mid | group:mid_healthy | Sample source:microglia genotype:WT | condition:low | group:low_healthy | Sample source:microglia genotype:APP | condition:mid | group:mid_disease | Sample source:microglia genotype:APP | condition:mid | group:mid_disease | Sample source:microglia genotype:APP | condition:low | group:low_disease | Sample source:microglia genotype:APP | condition:high | group:high_disease | Sample source:microglia genotype:WT | condition:mid | group:mid_healthy | Sample source:microglia genotype:WT | condition:mid | group:mid_healthy | Sample source:microglia genotype:APP | condition:mid | group:mid_disease | Sample source:microglia genotype:WT | condition:low | group:low_healthy | Sample source:microglia genotype:APP | condition:high | group:high_disease | Sample source:microglia genotype:WT | condition:low | group:low_healthy | Sample source:microglia genotype:NA | condition:NA | group:NA | Sample source:microglia genotype:WT | condition:mid | group:mid_healthy | Sample source:microglia genotype:WT | condition:low | group:low_healthy | Sample source:microglia genotype:APP | condition:low | group:low_disease | Sample source:microglia genotype:APP | condition:high | group:high_disease | Sample source:microglia genotype:APP | condition:high | group:high_disease | Sample source:microglia genotype:WT | condition:low | group:low_healthy | Sample source:microglia genotype:APP | condition:low | group:low_disease | Sample source:microglia genotype:WT | condition:low | group:low_healthy | Sample source:microglia genotype:APP | condition:mid | group:mid_disease | Sample source:microglia genotype:WT | condition:low | group:low_healthy | Sample source:microglia genotype:WT | condition:mid | group:mid_healthy | Sample source:microglia genotype:NA | condition:NA | group:NA | Sample source:microglia 13C3-Citric acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 13C4-Fumaric acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 13C4-Malic acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 13C4-Succinic acid 20.696272926743678 20.56548769821013 20.571484588381583 20.60595306722529 20.577617540913213 20.51421135263355 20.547209687416174 20.462518540143712 20.506008677119834 20.458784421344564 20.445906105607094 20.410826096914235 20.43848553113269 20.457611910420837 20.567061915240576 20.449200034610612 20.504149513710068 20.454897047883886 20.44352229956763 20.40123773607129 20.39130874866888 20.376138281188023 20.365287092295667 20.331311244460604 20.443176227409626 20.437544208820817 20.36565686146364 20.368737577144408 20.479123224002883 13C5-Oxoglutaric acid 19.758219507154585 19.70555627828419 19.729502641366466 19.72512586852337 19.702405379193117 19.69865233159412 19.545996543448485 19.635924634356318 19.63275348217121 19.498934065069676 19.68329348992736 19.652699616700815 19.515984648389978 19.479641768798647 19.405045728211824 19.640782344895523 19.52114971204955 19.480172798525686 19.558422981575095 19.557531350936625 19.48890971441678 19.531532975236782 19.541320179707295 19.565246201268273 19.43473686005052 19.436061475071917 19.528497690272143 19.430637199924195 19.536761183773873 13C6-Benzoic acid 21.244384241485037 21.126745786827975 21.21566042356713 21.21196377537931 21.22906608881889 21.278219949178126 21.090638032513844 21.209231086244973 21.237481349599364 21.079199394132882 21.32546241743073 21.273466475140026 21.177547778755155 21.094583082866297 21.017732486709843 21.20351988292637 21.162492971756404 21.155327332502846 21.161675728898043 21.171233068038635 21.115306065706303 21.214812290633528 21.146706780386133 21.221728226839467 21.11027100005672 21.091482090164334 21.19543060041183 21.218147207174095 21.155859230139885 13C8-Octanic acid 24.559944434946996 24.517408730554962 24.585789147734268 24.489152869663805 24.557102947456812 24.64629003618371 24.457801496552367 24.61672305038245 24.599571109999733 24.46979770922719 24.84447467400862 24.656994638232074 24.507629225912336 24.523111638846117 24.420174594029003 24.582709907507468 24.613994646829376 24.560306327111654 24.53717769917855 24.578608995184712 24.46365128076371 24.59046570153508 24.540385836111298 24.622421013624848 24.555562808434676 24.53989737954426 24.581221208447154 24.596252308153694 24.47762227788992 15N5-ADP 21.614297289512496 21.434733231514098 20.522726240071023 20.649786998769134 19.88579182 18.421240616687307 19.855439245656182 18.176646706816317 18.875145525915784 19.55215064110112 15.358328333638793 16.920700443264725 19.031238856175857 19.06555565593994 19.756209147841442 18.036365619502366 18.787432177181028 18.439799654436538 18.353977366289428 18.12430722621877 18.181114059436364 17.60225036541727 17.86204762820679 16.39703459317729 17.926629520837448 18.178789642813427 16.979444377657288 16.143096539585667 17.944409704860075 2-Hydroxyglutarate NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 2-Phosphoglyceric acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 3-Hydroxybutyric acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 3-Phosphoglyceric acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 6-Phosphogluconic acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Acetoacetic acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Adenosine monophosphate NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Adenosine triphosphate NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Adenosine triphosphate-d4 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA ADP NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Alanine NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Alanine-d4 21.615386579176345 21.27622048283794 21.656431846710568 21.547741370100518 21.648713229977812 21.97308631435992 21.442792131695533 21.935266135589792 21.73913465522812 21.43515116015182 22.18160595847872 22.014364793239302 21.526889371072265 21.430094323967623 20.59720958430099 21.832851543070774 21.534066876559052 21.62350982134825 21.604383542769444 21.683808356099707 21.48600724092819 21.834045135247816 21.64571682126857 21.908180754435133 21.614477381970925 21.348451366333965 21.91700418286355 21.970221203005195 21.556290459323286 Arginine NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Arginine-d4C13 25.94338437254793 25.85023156938372 25.95409136261772 25.92129832902276 25.912878670198996 25.931865684628818 25.807916446307132 26.06439717156318 25.918700515873667 25.858296820709736 25.99872426018753 25.97110928781179 25.810703671390772 25.767757868155716 25.44607868801695 25.862951505057488 25.790734045902557 25.75358506553661 25.814678040007408 25.70313965400282 25.689125967323246 25.792966576281813 25.809914341665163 25.864986104675005 25.74173047 25.59392051363478 25.821239847282925 25.82704939 25.668026200767255 Asparagine NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Aspartic acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Aspartic acid-d3 24.56448049943304 24.411441222880207 24.440367358659003 24.361417782094446 24.406289724112895 24.371282262520637 24.24401941991085 24.357755072100435 24.260971655404735 24.162130319331133 24.361873935421045 24.183967158442965 24.15691755 24.184094493245993 24.033897995393957 24.27843148057162 24.164510243859965 24.072013463344597 24.101887652659148 24.134456746741485 24.07755511880683 24.156201572752323 24.077080669977143 24.167677313538636 24.092425468297133 24.033743465691618 24.203110478854757 24.029661113511477 23.94574908033826 cis-Aconitic acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Sum of Citric acid and Isocitric acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Dihydroxyacetone phosphate NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Erythose 4-phosphate NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Fructose 1,6-bisphosphate 16.94931551 17.073847008686805 17.03031340567249 17.12977934474663 16.95040562060058 16.21279059081403 17.139546878982976 16.279081683157088 16.703988320625136 17.078328783325755 15.750423337171632 16.11362468808342 17.261831860742934 17.22526055668612 17.33575803 16.46591226567112 17.28644811279494 16.807688133186332 16.903123259875844 16.623047672366713 16.834526475150064 16.574908047985005 16.707226642105958 15.923087376208967 16.971174461083706 17.17440309086494 16.518159420627686 16.127103011275405 17.264404128474993 Fructose 6-phosphate NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Fumaric acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Glucose 1-phosphate NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Glucose 6-phosphate 16.695414622562815 16.963327725154468 16.60193802860686 16.88833176413429 16.092266040240638 15.264891177786424 17.032439334884153 16.187168027020586 16.099426656112435 15.97847753139458 14.781359287745575 16.730411680913765 16.05141856490009 15.537329642145874 16.23516130664381 17.057965332886628 16.730476594034272 17.27583924303307 15.637673327333621 15.888527495393689 16.423351801615333 16.002153700883145 15.87752128614099 15.138507615561265 16.117453384472803 15.907860000049144 16.57597978397044 14.762959378555447 16.33766975000846 Glutamic acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Glutamic acid-d3 22.68369998136673 22.608184089738465 22.70631943810525 22.634042628577635 22.64296368786774 22.60401898524518 22.479548250361233 22.583578304122934 22.50766014342594 22.48299584219232 22.67355666118105 22.62468155307527 22.43106162008862 22.492871942188366 22.255020303914016 22.530454606941653 22.486253438247125 22.221635413066057 22.427996092053455 22.42834237644278 22.337045030054338 22.454277432763412 22.444048258004127 22.418772630754795 22.406121449063516 22.373164889369516 22.49971498944773 22.416708491217996 22.379260050625152 Glutamine NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Glyceraldehyde 3-phosphate NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Lactic acid 17.195378230638894 15.774453094754179 16.74538879505487 16.122115401832346 16.032770410189972 15.52989571 16.377083481121588 17.243197394099997 15.708553696193125 14.964775560557019 17.40883281276612 17.26204935543534 15.467535224746054 15.648330037246607 15.596573079723203 19.07084460559871 17.07095712676364 16.862191326862245 15.110380766950188 14.727489022884454 17.296038193116445 16.10461583884974 15.084624844198098 15.277055275607275 16.571194127503862 16.203789379921044 15.820271626018814 16.958292795853907 17.03673796177161 Malic acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NAD NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NADH NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NADP NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NADPH NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Oxalacetic acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Oxoglutaric acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Phosphocreatine 16.12617234991907 16.87394690190043 16.785514466675536 15.972642568235722 16.17522474583265 13.982386237007377 17.255193660993736 16.26042718550107 15.70122004169764 16.33153261414818 13.32151416952848 17.259156501100357 15.628290720405769 15.587918172444539 16.236490132310454 15.85672981440694 16.16137611155079 17.015373496631362 15.549396341314733 15.746357281592433 15.852108824348452 16.110275287875943 15.15377653684824 16.08364528382903 13.936243374121855 16.395088414044288 15.81162671 14.09122615157264 15.80918798784736 Phosphoenolpyruvic acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Pyruvic acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Ribose 5-phosphate NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Ribulose 5-phosphate NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Sedoheptulose 7-phosphate 15.684096057185549 15.764049457348511 15.543463582032592 15.488660546651268 15.554463014277538 14.055593940495486 15.66960856310152 14.812106799349264 15.187963993005194 16.046873045936348 13.655506412633837 15.361074577479886 16.294562794115034 14.417238780546827 14.897173148807683 15.39243470317673 15.303773050612877 15.606885777732597 14.095183817368612 14.485740837694976 16.207154250972962 14.391967620348842 14.769241111454356 14.449427347177355 14.190992830160443 14.478490089745026 15.037922821239622 14.041550733726963 15.20495227495096 Serine NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Succinic acid 19.081376235834163 19.275626447657352 19.38011169590339 19.13389326362427 19.410767158098977 19.010072074351207 19.26240447868939 18.932402049449106 19.071856679844206 18.856869804140654 18.901376068142437 18.801602644992133 18.937999085875962 19.07632531202259 18.728553453922004 19.098369768981588 18.811128617640218 19.04669016307844 19.21816219058364 18.67631903397029 19.341565506739933 18.785321592587724 18.65298594608943 18.69705016730237 18.841941968448825 18.67175992036619 18.695966146780155 18.89221732658571 18.77438311337504 U-13C10,U-15N5-Adenosine monophosphate NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA U-13C3-Pyruvic acid 17.089736248739968 16.926165551215057 17.18011515 17.137269990363855 17.13871728 17.382602073646485 16.75578210505539 17.19360679820826 17.133725101874116 16.918344917797636 17.496910829732784 17.192974831983644 16.98649550187652 16.774085087217134 16.638741831572545 16.93971407375806 16.87476180908621 16.945095534696048 16.941034283054613 16.959833522182016 16.83513056 17.104683952637735 16.968128894401644 17.235494854519278 16.814811452945026 16.80277876239668 16.979108351049806 17.066357298342908 16.73760370523315 U-13C6-Glucose 6-phosphate 23.114845958824638 23.102070882825195 22.75768303 23.027736376017977 22.867167627733956 22.447236993482385 23.106844736899163 22.466802556298305 22.879870913794793 23.324380750168665 21.89485200492282 22.372293978071152 23.392868370602585 23.440491702285385 23.497974512697205 23.052436120571677 23.442426321403886 23.25658314637172 23.25682877596059 23.14966719 23.194762976133028 23.108822195513834 23.334854473271555 22.96748910367867 23.362165405246948 23.388962103767593 23.089631121196376 22.76564368041822 23.271849434498037 Xylulose 5-phosphate NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Fructose 1-phosphate NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name internal standard Expected RT precursor (m/z) product (m/z) 13C3-Citric acid NA 3.77 194.2 113 13C4-Fumaric acid NA 3.28 119.2 74.3 13C4-Malic acid NA 3.26 137.2 119.2 13C4-Succinic acid NA 3.07 121.3 76 13C5-Oxoglutaric acid NA 2.97 150.2 105.3 13C6-Benzoic acid NA 0.51 127 83 13C8-Octanic acid NA 1.04 151.1 151.1 15N5-ADP NA 3.55 431.1 79 2-Hydroxyglutarate 13C4-Succinic acid 2.97 145 101 2-Phosphoglyceric acid 13C4-Succinic acid 3.66 185.5 97.1 3-Hydroxybutyric acid 13C4-Succinic acid 0.94 103.2 59.2 3-Phosphoglyceric acid 13C4-Succinic acid 3.67 185.5 97 6-Phosphogluconic acid 13C4-Succinic acid 3.77 275 97 Acetoacetic acid 13C4-Succinic acid 0.31 101 101 Adenosine monophosphate U-13C10,U-15N5-Adenosine monophosphate 3.27 346 79 Adenosine triphosphate Adenosine triphosphate-d4 3.71 506.1 158.9 Adenosine triphosphate-d4 NA 3.7 510 158.9 ADP 15N5-ADP 3.53 426.1 79 Alanine Alanine-d4 2.18 88 88 Alanine-d4 NA 2 92 92 Arginine Arginine-d4C13 3.95 173 131 Arginine-d4C13 NA 3.96 178 136 Asparagine 13C4-Succinic acid 2.48 131 114 Aspartic acid Aspartic acid-d3 3.05 132 88 Aspartic acid-d3 NA 3.06 135 91 cis-Aconitic acid 13C4-Succinic acid 3.29 173 129 Sum of Citric acid and Isocitric acid 13C4-Succinic acid 3.77 191.2 111 Dihydroxyacetone phosphate 13C4-Succinic acid 3.35 169.1 79 Erythose 4-phosphate 13C4-Succinic acid 3.65 199 97 Fructose 1,6-bisphosphate 13C4-Succinic acid 3.48 339.1 79 Fructose 6-phosphate 13C4-Succinic acid 3.5 259 97.1 Fumaric acid 13C4-Succinic acid 6.34 115.2 71.3 Glucose 1-phosphate 13C5-Oxoglutaric acid 3.62 259 241 Glucose 6-phosphate 13C8-Octanic acid 3.64 259 97 Glutamic acid 13C4-Succinic acid 3.06 146 128 Glutamic acid-d3 NA 3.02 149 131 Glutamine 13C4-Succinic acid 2.46 145 127 Glyceraldehyde 3-phosphate 13C4-Succinic acid 3.35 169.1 97 Lactic acid 13C4-Succinic acid 0.87 89.2 43.2 Malic acid 13C4-Succinic acid 3.46 133.2 115.2 NAD 13C4-Succinic acid 3.22 662.1 540.3 NADH 13C4-Succinic acid 3.03 664.1 408.1 NADP 13C4-Succinic acid 3.92 742.1 620 NADPH 13C4-Succinic acid 3.75 744.1 408 Oxalacetic acid 13C5-Oxoglutaric acid 3.63 131 131 Oxoglutaric acid 13C5-Oxoglutaric acid 2.96 145.2 101.3 Phosphocreatine 13C4-Succinic acid 3.68 210 79 Phosphoenolpyruvic acid 13C4-Succinic acid 3.78 167 79 Pyruvic acid 13C8-Octanic acid 0.5 87.2 43.2 Ribose 5-phosphate 13C4-Succinic acid 3.35 229 97 Ribulose 5-phosphate 13C4-Succinic acid 3.42 229 97.1 Sedoheptulose 7-phosphate 13C4-Succinic acid 3.55 289 97 Serine 13C4-Succinic acid 2.44 104 74 Succinic acid 13C4-Succinic acid 3.07 117.2 73.2 U-13C10,U-15N5-Adenosine monophosphate NA 3.28 361 79 U-13C3-Pyruvic acid NA 0.49 90.2 45.2 U-13C6-Glucose 6-phosphate NA 3.71 265 97 Xylulose 5-phosphate 13C4-Succinic acid 3.34 229.1 97 Fructose 1-phosphate 13C4-Succinic acid 3.63 259.1 79 METABOLITES_END #END