#METABOLOMICS WORKBENCH nmakdissi_20241203_093051 DATATRACK_ID:5420 STUDY_ID:ST003615 ANALYSIS_ID:AN005940 PROJECT_ID:PR002234 VERSION 1 CREATED_ON 02-06-2025 #PROJECT PR:PROJECT_TITLE Kupffer cells control neonatal hepatic glucose metabolism via Igf1 signaling PR:PROJECT_TYPE lipidomics analysis PR:PROJECT_SUMMARY During perinatal development, liver metabolism is tightly regulated to ensure PR:PROJECT_SUMMARY energy supply for the newborn. Before birth, glycogen is stored in hepatocytes PR:PROJECT_SUMMARY and later metabolized to glucose, meeting the energy demands of the neonate. PR:PROJECT_SUMMARY Shortly after birth, lipogenesis begins, driven by the transcriptional PR:PROJECT_SUMMARY activation of enzymes involved in fatty acid oxidation. These processes are PR:PROJECT_SUMMARY thought to be largely regulated by systemic insulin and glucagon levels. PR:PROJECT_SUMMARY However, the role of liver-derived local factors in neonatal hepatocyte PR:PROJECT_SUMMARY metabolism remains unexplored. Kupffer cells (KCs), the liverâs resident PR:PROJECT_SUMMARY macrophages, colonize the fetal liver early in embryogenesis and support liver PR:PROJECT_SUMMARY metabolism in adulthood. Yet, whether KCs influence neonatal hepatocyte PR:PROJECT_SUMMARY metabolism is unknown. Here, using conditional knockout mouse models targeting PR:PROJECT_SUMMARY macrophages, we demonstrate that yolk sac-derived KCs play a critical role in PR:PROJECT_SUMMARY hepatocyte glycogen storage and function by regulating the TCA cycle - a role PR:PROJECT_SUMMARY that monocyte-derived KC-like cells cannot substitute. Newborn pups lacking KCs PR:PROJECT_SUMMARY mobilize glycogen more rapidly, a process regulated by insulin-like growth PR:PROJECT_SUMMARY factor 1 (Igf1) production. Our findings reveal that macrophages are a major PR:PROJECT_SUMMARY source of Igf1 at birth and that local Igf1 production by KCs is essential for PR:PROJECT_SUMMARY balanced hepatocyte metabolism. PR:INSTITUTE University of Bonn PR:DEPARTMENT Developmental Biology of the Immune System, The Life & Medical Sciences PR:DEPARTMENT Institute (LIMES) PR:LABORATORY Mass Lab PR:LAST_NAME Makdissi PR:FIRST_NAME Nikola PR:ADDRESS Carl Troll straÃe 31, Bonn PR:EMAIL nmakdissi@uni-bonn.de PR:PHONE 02 28 / 73 - 6 2794 PR:DOI http://dx.doi.org/10.21228/M8VJ9K #STUDY ST:STUDY_TITLE Kupffer cells control neonatal hepatic glucose metabolism via Igf1 signaling - ST:STUDY_TITLE C13 glucose traced metabolites analysis of postnatal day 0 livers after ST:STUDY_TITLE macrophages depletion ST:STUDY_TYPE metabolites analysis of P0 livers from mice ST:STUDY_SUMMARY During perinatal development, liver metabolism is tightly regulated to ensure ST:STUDY_SUMMARY energy supply for the newborn. Before birth, glycogen is stored in hepatocytes ST:STUDY_SUMMARY and later metabolized to glucose, meeting the energy demands of the neonate. ST:STUDY_SUMMARY Shortly after birth, lipogenesis begins, driven by the transcriptional ST:STUDY_SUMMARY activation of enzymes involved in fatty acid oxidation. These processes are ST:STUDY_SUMMARY thought to be largely regulated by systemic insulin and glucagon levels. ST:STUDY_SUMMARY However, the role of liver-derived local factors in neonatal hepatocyte ST:STUDY_SUMMARY metabolism remains unexplored. Kupffer cells (KCs), the liverâs resident ST:STUDY_SUMMARY macrophages, colonize the fetal liver early in embryogenesis and support liver ST:STUDY_SUMMARY metabolism in adulthood. Yet, whether KCs influence neonatal hepatocyte ST:STUDY_SUMMARY metabolism is unknown. Here, using conditional knockout mouse models targeting ST:STUDY_SUMMARY macrophages, we demonstrate that yolk sac-derived KCs play a critical role in ST:STUDY_SUMMARY hepatocyte glycogen storage and function by regulating the TCA cycle - a role ST:STUDY_SUMMARY that monocyte-derived KC-like cells cannot substitute. n this experiment, ST:STUDY_SUMMARY C13-labeled glucose is administered to track glucose metabolism. The labeled ST:STUDY_SUMMARY glucose is metabolized, and its incorporation into downstream metabolites is ST:STUDY_SUMMARY analyzed using techniques such as mass spectrometry . This approach enables the ST:STUDY_SUMMARY identification and quantification of metabolic pathways involving glucose, such ST:STUDY_SUMMARY as glycolysis, gluconeogenesis, and the TCA cycle, providing insights into ST:STUDY_SUMMARY glucose utilization and related metabolic processes. ST:INSTITUTE University of Bonn ST:DEPARTMENT Developmental Biology of the Immune System, The Life & Medical Sciences ST:DEPARTMENT Institute (LIMES) ST:LABORATORY Mass Lab ST:LAST_NAME Makdissi ST:FIRST_NAME Nikola ST:ADDRESS Carl Troll straÃe 31 ST:EMAIL nmakdissi@uni-bonn.de ST:PHONE 02 28 / 73 - 6 2794 ST:SUBMIT_DATE 2024-12-03 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - 24_KO1 Sample source:liver | Genotype:KO replicate=replicate1; RAW_FILE_NAME(row data file name)=fni_Nikola_24.cdf SUBJECT_SAMPLE_FACTORS - 25_KO1 Sample source:liver | Genotype:KO replicate=replicate2; RAW_FILE_NAME(row data file name)=fni_Nikola_25.cdf SUBJECT_SAMPLE_FACTORS - 26_KO1 Sample source:liver | Genotype:KO replicate=replicate3; RAW_FILE_NAME(row data file name)=fni_Nikola_26.cdf SUBJECT_SAMPLE_FACTORS - 27_KO2 Sample source:liver | Genotype:KO replicate=replicate1; RAW_FILE_NAME(row data file name)=fni_Nikola_27.cdf SUBJECT_SAMPLE_FACTORS - 28_KO2 Sample source:liver | Genotype:KO replicate=replicate2; RAW_FILE_NAME(row data file name)=fni_Nikola_28.cdf SUBJECT_SAMPLE_FACTORS - 29_KO2 Sample source:liver | Genotype:KO replicate=replicate3; RAW_FILE_NAME(row data file name)=fni_Nikola_29.cdf SUBJECT_SAMPLE_FACTORS - 50_KO3 Sample source:liver | Genotype:KO replicate=replicate1; RAW_FILE_NAME(row data file name)=fni_Nikola_50.cdf SUBJECT_SAMPLE_FACTORS - 51_KO3 Sample source:liver | Genotype:KO replicate=replicate2; RAW_FILE_NAME(row data file name)=fni_Nikola_51.cdf SUBJECT_SAMPLE_FACTORS - 52_KO3 Sample source:liver | Genotype:KO replicate=replicate3; RAW_FILE_NAME(row data file name)=fni_Nikola_52.cdf SUBJECT_SAMPLE_FACTORS - 59_KO4 Sample source:liver | Genotype:KO replicate=replicate1; RAW_FILE_NAME(row data file name)=fni_Nikola_59.cdf SUBJECT_SAMPLE_FACTORS - 60_KO4 Sample source:liver | Genotype:KO replicate=replicate2; RAW_FILE_NAME(row data file name)=fni_Nikola_60.cdf SUBJECT_SAMPLE_FACTORS - 61_KO4 Sample source:liver | Genotype:KO replicate=replicate3; RAW_FILE_NAME(row data file name)=fni_Nikola_61.cdf SUBJECT_SAMPLE_FACTORS - 18_WT1 Sample source:liver | Genotype:WT replicate=replicate1; RAW_FILE_NAME(row data file name)=fni_Nikola_18.cdf SUBJECT_SAMPLE_FACTORS - 19_WT1 Sample source:liver | Genotype:WT replicate=replicate2; RAW_FILE_NAME(row data file name)=fni_Nikola_19.cdf SUBJECT_SAMPLE_FACTORS - 20_WT1 Sample source:liver | Genotype:WT replicate=replicate3; RAW_FILE_NAME(row data file name)=fni_Nikola_20.cdf SUBJECT_SAMPLE_FACTORS - 21_WT2 Sample source:liver | Genotype:WT replicate=replicate1; RAW_FILE_NAME(row data file name)=fni_Nikola_21.cdf SUBJECT_SAMPLE_FACTORS - 22_WT2 Sample source:liver | Genotype:WT replicate=replicate2; RAW_FILE_NAME(row data file name)=fni_Nikola_22.cdf SUBJECT_SAMPLE_FACTORS - 23_WT2 Sample source:liver | Genotype:WT replicate=replicate3; RAW_FILE_NAME(row data file name)=fni_Nikola_23.cdf SUBJECT_SAMPLE_FACTORS - 53_WT3 Sample source:liver | Genotype:WT replicate=replicate1; RAW_FILE_NAME(row data file name)=fni_Nikola_53.cdf SUBJECT_SAMPLE_FACTORS - 54_WT3 Sample source:liver | Genotype:WT replicate=replicate2; RAW_FILE_NAME(row data file name)=fni_Nikola_54.cdf SUBJECT_SAMPLE_FACTORS - 55_WT3 Sample source:liver | Genotype:WT replicate=replicate3; RAW_FILE_NAME(row data file name)=fni_Nikola_55.cdf SUBJECT_SAMPLE_FACTORS - 62_WT4 Sample source:liver | Genotype:WT replicate=replicate1; RAW_FILE_NAME(row data file name)=fni_Nikola_62.cdf SUBJECT_SAMPLE_FACTORS - 63_WT4 Sample source:liver | Genotype:WT replicate=replicate2; RAW_FILE_NAME(row data file name)=fni_Nikola_63.cdf SUBJECT_SAMPLE_FACTORS - 66_WT4 Sample source:liver | Genotype:WT replicate=replicate3; RAW_FILE_NAME(row data file name)=fni_Nikola_66.cdf #COLLECTION CO:COLLECTION_SUMMARY The pups were sacrificed directly after birth by decapitation, and the liver was CO:COLLECTION_SUMMARY collected in ice-cold culture buffer (Williams Medium supplemented with 10% FCS, CO:COLLECTION_SUMMARY 1 % L-glutamine, and 1% PenStrep)). Between 2-3 mg per replicate of the biggest CO:COLLECTION_SUMMARY liver lobe were carefully dissected and placed in a 48-well plate on ice-cold CO:COLLECTION_SUMMARY culture buffer. All the following steps were performed in a sterile hood. Each CO:COLLECTION_SUMMARY piece was carefully placed on a porous cell culture insert (0.4 µm pore size), CO:COLLECTION_SUMMARY which was then placed in a well of a 24-well plate containing 400 µL prewarmed CO:COLLECTION_SUMMARY culture buffer. The plate was incubated for 2 h at 37°C, 5 % CO2. This step was CO:COLLECTION_SUMMARY necessary to allow for the simultaneous genotyping of the mice to ensure that CO:COLLECTION_SUMMARY only WTSpi and KOSpi were used in the tracing experiment, not HETSpi. Once the CO:COLLECTION_SUMMARY genotypes were determined, the respective inlets were transferred to a 24-well CO:COLLECTION_SUMMARY plate containing 400 µL pre-warmed tracer medium (Williams Medium w/o Glucose CO:COLLECTION_SUMMARY supplemented with 10% FCS, 1 % L-glutamine, and 25 mM C13 labeled Glucose. The CO:COLLECTION_SUMMARY samples were incubated for 4 h at 37°C, 5 % CO2. After incubation 100 µL of CO:COLLECTION_SUMMARY medium was collected and snap-frozen. The liver tissue piece was briefly washed CO:COLLECTION_SUMMARY with 0.9 % NaCl and then placed in 250 µL of -20°C cold Methanol. 250 µL of CO:COLLECTION_SUMMARY pre-cooled MS-grade ddH2O containing 1 µg/mL D6 glutaric acid was added and the CO:COLLECTION_SUMMARY tissue homogenized. The homogenate was transferred to a fresh tube and stored at CO:COLLECTION_SUMMARY -80°C until further processing. CO:SAMPLE_TYPE Liver #TREATMENT TR:TREATMENT_SUMMARY No treatment. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Metabolites were derivatized using a Gerstel MPS with 15 μL of 2% (w/v) SP:SAMPLEPREP_SUMMARY methoxyamine hydrochloride (Thermo Scientific) in pyridine and 15 μl SP:SAMPLEPREP_SUMMARY N-tertbutyldimethylsilyl-N-methyltrifluoroacetamide (MTBSTFA) with 1% SP:SAMPLEPREP_SUMMARY tert-butyldimethylchlorosilane (Regis Technologies). #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Derivatives were measured by GC/MS with a 30 m DB-35MS + 5 m Duraguard capillary CH:CHROMATOGRAPHY_SUMMARY column (0.25 mm inner diameter, 0.25 µm film thickness) equiped in an Agilent CH:CHROMATOGRAPHY_SUMMARY 7890B gas chromatograph (GC) connected to an Agilent 5977A mass spectrometer CH:CHROMATOGRAPHY_SUMMARY (MS). The GC oven temperature was held at 80°C for 6âmin and subsequently CH:CHROMATOGRAPHY_SUMMARY increased at 6°C per min until reaching 280°C where it was held for 10âmin. CH:INSTRUMENT_NAME Agilent 7890B CH:COLUMN_NAME Agilent DB5-MS (30m x 0.25mm, 0.25um) CH:COLUMN_TEMPERATURE The oven temperature was initially set at 80°C for 6âminutes, ramped at 6°C CH:COLUMN_TEMPERATURE per minute to 280°C, and held at 280°C for 10âminutes CH:FLOW_GRADIENT NA CH:FLOW_RATE 1 mL/min CH:SOLVENT_A NA CH:SOLVENT_B NA CH:CHROMATOGRAPHY_TYPE GC #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 5977A MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:MS_COMMENTS The quadropole was held at 150°C. The MS source operated under electron impact MS:MS_COMMENTS ionization mode at 70 eV and was held at 230°C. Full scan (70â800âm/z, 3.9 MS:MS_COMMENTS scans per second) as well as targeted ion chromatogram measurements were MS:MS_COMMENTS conducted for pyruvate (174, 175, 176, 177, 178, 179; 10 scans per second), MS:MS_COMMENTS lactate (261, 262, 263, 264, 265, 266, 267; 10 scans per second), alanine (260, MS:MS_COMMENTS 261, 262, 263, 264, 265; 10 scans per second), citrate (591, 592, 593, 594, 595, MS:MS_COMMENTS 596, 597, 598, 599, 600; 10 scans per second), malate (419, 420, 421, 422, 423, MS:MS_COMMENTS 424, 425, 426; 10 scans per second) and aspartate (481, 482, 483, 484, 485, 486, MS:MS_COMMENTS 487; 10 scans per second) All chromatograms were analyzed with MS:MS_COMMENTS MetaboliteDetector [REF: Hiller K, Hangebrauk J, Jäger C, Spura J, Schreiber K, MS:MS_COMMENTS Schomburg D. MetaboliteDetector: comprehensive analysis tool for targeted and MS:MS_COMMENTS nontargeted GC/MS based metabolome analysis. Anal Chem. 2009 May MS:MS_COMMENTS 1;81(9):3429-39. doi: 10.1021/ac802689c. PMID: 19358599.]. MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Signal intensity normalized by IS for the metabolite signal/c13 carbon enrichment % MS_METABOLITE_DATA_START Samples 24_KO1 25_KO1 26_KO1 27_KO2 28_KO2 29_KO2 50_KO3 51_KO3 52_KO3 59_KO4 60_KO4 61_KO4 18_WT1 19_WT1 20_WT1 21_WT2 22_WT2 23_WT2 53_WT3 54_WT3 55_WT3 62_WT4 63_WT4 66_WT4 Factors Sample source:liver | Genotype:KO Sample source:liver | Genotype:KO Sample source:liver | Genotype:KO Sample source:liver | Genotype:KO Sample source:liver | Genotype:KO Sample source:liver | Genotype:KO Sample source:liver | Genotype:KO Sample source:liver | Genotype:KO Sample source:liver | Genotype:KO Sample source:liver | Genotype:KO Sample source:liver | Genotype:KO Sample source:liver | Genotype:KO Sample source:liver | Genotype:WT Sample source:liver | Genotype:WT Sample source:liver | Genotype:WT Sample source:liver | Genotype:WT Sample source:liver | Genotype:WT Sample source:liver | Genotype:WT Sample source:liver | Genotype:WT Sample source:liver | Genotype:WT Sample source:liver | Genotype:WT Sample source:liver | Genotype:WT Sample source:liver | Genotype:WT Sample source:liver | Genotype:WT Alanine_total 0.0248 0.0240 0.0182 0.0216 0.0225 0.0193 0.0186 0.0303 0.0179 0.0204 0.0243 0.0234 0.0180 0.0268 0.0179 0.0211 0.0281 0.0171 0.0228 0.0214 0.0155 0.0202 0.0131 0.0217 Aspartate_total 0.8703 0.7100 1.1252 1.1617 1.4820 1.2913 1.4397 0.9229 1.3318 0.8159 1.2926 0.7465 0.8080 1.6321 0.8025 0.7877 0.6935 0.9131 1.9359 1.4754 0.8529 0.9950 1.5904 1.4806 Citrate_total 0.5080 0.5561 0.5279 0.5403 0.4631 0.4986 0.5057 0.5116 0.5144 0.5267 0.5206 0.5315 0.5721 0.4954 0.4309 0.5655 0.4987 0.5165 0.5655 0.4898 0.5106 0.5387 0.5461 0.4877 Lactate_total 12.0704 10.4378 10.6633 10.8889 10.7813 13.2998 14.3095 13.8050 14.2657 9.2289 10.4637 7.0253 6.6553 4.7383 6.9239 8.7587 4.0443 7.5498 6.0467 4.1825 4.6167 5.6722 4.1919 4.6546 Malate_total 1.6926 0.8430 0.9229 1.5725 0.6469 1.5015 1.1382 0.7251 1.2416 1.8961 0.8587 0.9445 1.2223 0.9909 0.9249 0.8223 0.9401 1.4691 0.9791 1.4042 1.5566 1.4787 1.2605 1.1017 Pyruvate_total 0.9460 1.0321 1.3484 0.9923 0.9412 1.0171 0.8821 0.8703 0.8131 0.9350 0.9909 0.9236 0.1335 0.1545 0.1796 0.3638 0.1365 0.2188 0.2487 0.2862 0.1144 0.1444 0.1167 0.1475 %traced_Alanine_260 (M0) 0.5080 0.4923 0.5091 0.4712 0.5100 0.5240 0.5313 0.5067 0.5429 0.5253 0.5306 0.5009 0.4698 0.4628 0.5150 0.4958 0.5138 0.5033 0.5103 0.5388 0.4477 0.5163 0.5379 0.5240 %traced_Alanine_261 (M1) 0.0462 0.0409 0.0509 0.0519 0.0352 0.0243 0.0170 0.0237 0.0350 0.0872 0.0123 0.0752 0.0498 0.0434 0.0479 0.0334 0.0299 0.0370 0.0527 0.0308 0.0280 0.0301 0.0118 0.0089 %traced_Alanine_262 (M2) 0.0757 0.0761 0.0730 0.0344 0.0644 0.0538 0.0031 0.0582 0.0018 0.0371 0.0810 0.0775 0.0824 0.0998 0.0281 0.0744 0.0711 0.0591 0.0641 0.0219 0.0630 0.0597 0.0293 0.0725 %traced_Alanine_263 (M3) 0.4031 0.3939 0.4002 0.4097 0.3925 0.3996 0.3803 0.3936 0.3716 0.4158 0.4017 0.3654 0.3992 0.4219 0.4154 0.4022 0.3939 0.3957 0.3923 0.3885 0.3919 0.3973 0.4235 0.3881 %traced_Aspartate_418 (M0) 0.5880 0.6027 0.5835 0.5992 0.6313 0.5958 0.6364 0.6285 0.6173 0.6479 0.5965 0.6423 0.6478 0.6080 0.6538 0.6345 0.5889 0.5901 0.6576 0.6091 0.6325 0.6272 0.6496 0.5978 %traced_Aspartate_419 (M1) 0.0314 0.0161 0.0651 0.0182 0.0199 0.0547 0.0872 0.0818 0.0635 0.0900 0.0742 0.0708 0.0995 0.0402 0.0801 0.0480 0.0674 0.0575 0.0338 0.0704 0.0298 0.0830 0.0755 0.0862 %traced_Aspartate_420 (M2) 0.2576 0.2353 0.2119 0.1924 0.2062 0.2003 0.2341 0.1825 0.2093 0.2010 0.2089 0.2329 0.2024 0.2068 0.2365 0.2307 0.2021 0.2144 0.2483 0.1837 0.2102 0.2246 0.1540 0.2148 %traced_Aspartate_421 (M3) 0.1343 0.0936 0.0761 0.1077 0.0919 0.1167 0.1387 0.0924 0.1273 0.1155 0.1064 0.1168 0.0537 0.0763 0.1191 0.0775 0.0938 0.0871 0.1535 0.1184 0.0950 0.1249 0.1166 0.1106 %traced_Aspartate_422 (M4) 0.0080 0.0092 0.0077 0.0175 0.0111 0.0082 0.0121 0.0195 0.0111 0.0063 0.0066 0.0143 0.0220 0.0077 0.0084 0.0108 0.0086 0.0070 0.0062 0.0040 0.0072 0.0081 0.0059 0.0075 %traced_Citrate_591 (M0) 0.6707 0.7258 0.6621 0.7290 0.6931 0.6775 0.7501 0.7442 0.7111 0.7258 0.6931 0.7069 0.6581 0.6669 0.6890 0.6501 0.6442 0.6111 0.6538 0.6581 0.6333 0.5290 0.5775 0.5581 %traced_Citrate_592 (M1) 0.0102 0.0408 0.0211 0.0134 0.0127 0.0120 0.0286 0.0301 0.0400 0.0408 0.0127 0.0379 0.0109 0.0379 0.0126 0.0286 0.0301 0.0400 0.0364 0.0393 0.0623 0.0336 0.0120 0.0109 %traced_Citrate_593 (M2) 0.1606 0.1722 0.2223 0.2003 0.2179 0.2537 0.2858 0.2675 0.2801 0.1722 0.2179 0.2145 0.2964 0.2545 0.2562 0.2858 0.2675 0.2801 0.2869 0.2964 0.3008 0.3003 0.2537 0.2964 %traced_Citrate_594 (M3) 0.0555 0.0138 0.0411 0.0415 0.0200 0.0197 0.0585 0.0284 0.0196 0.0138 0.0200 0.0162 0.0844 0.0618 0.0880 0.0585 0.0284 0.0196 0.1104 0.0844 0.1232 0.0915 0.0972 0.0844 %traced_Citrate_595 (M4) 0.0126 0.0198 0.0333 0.0294 0.0414 0.0209 0.0764 0.0810 0.0867 0.0978 0.0914 0.0904 0.0397 0.0504 0.0409 0.0764 0.0810 0.0367 0.0292 0.0397 0.0686 0.0937 0.0209 0.0397 %traced_Citrate_596 (M5) 0.0230 0.0780 0.0137 0.0856 0.0172 0.0676 0.0308 0.0294 0.0211 0.0780 0.0172 0.0215 0.0120 0.0215 0.0388 0.0308 0.0294 0.0211 0.0703 0.0120 0.0250 0.0856 0.0676 0.0120 %traced_Citrate_597 (M6) 0.0126 0.0097 0.0161 0.0208 0.0013 0.0045 0.0020 0.0398 0.0853 0.0971 0.0013 0.0320 0.0009 0.0320 0.0313 0.0020 0.0398 0.0853 0.0016 0.0009 0.0118 0.0208 0.0452 0.0009 %traced_lactate_261 (M0) 0.3172 0.3173 0.3149 0.3296 0.3665 0.3689 0.2920 0.3195 0.3445 0.3914 0.3425 0.3591 0.4499 0.3999 0.4182 0.3774 0.4079 0.3478 0.4171 0.4352 0.4824 0.4199 0.4021 0.3948 %traced_lactate_262 (M1) 0.0525 0.0491 0.0551 0.0539 0.0516 0.0589 0.0337 0.0334 0.0378 0.0359 0.0362 0.0457 0.0386 0.0402 0.0314 0.0553 0.0543 0.0517 0.0466 0.0450 0.0463 0.0320 0.0406 0.0391 %traced_lactate_263 (M2) 0.0801 0.0850 0.0322 0.0854 0.0896 0.0839 0.0763 0.0785 0.0803 0.0731 0.0816 0.0849 0.0728 0.0754 0.0644 0.0921 0.0869 0.0893 0.0881 0.0807 0.0690 0.0685 0.0849 0.0784 %traced_lactate_264 (M3) 0.5531 0.5519 0.5803 0.5340 0.4956 0.4908 0.5999 0.5693 0.5403 0.5022 0.5421 0.5123 0.4419 0.4873 0.4893 0.4775 0.4538 0.5139 0.4511 0.4428 0.4051 0.4819 0.4752 0.4904 %traced_malate_419 (M0) 0.6779 0.5957 0.6477 0.6110 0.5879 0.5788 0.5734 0.6297 0.6534 0.6177 0.6154 0.6353 0.6454 0.6973 0.6873 0.6779 0.5957 0.6477 0.5579 0.6036 0.5816 0.5960 0.6097 0.5842 %traced_malate_420 (M1) 0.0725 0.0825 0.0855 0.0332 0.1144 0.0626 0.0640 0.0538 0.0631 0.0607 0.0613 0.0621 0.0847 0.0750 0.0610 0.0725 0.0825 0.0855 0.1095 0.1045 0.0954 0.0049 0.0505 0.0624 %traced_malate_421 (M2) 0.1264 0.1506 0.1341 0.1943 0.1546 0.1353 0.2277 0.2376 0.1591 0.1845 0.2045 0.1697 0.1484 0.1553 0.1457 0.1264 0.1506 0.1341 0.1482 0.1194 0.2217 0.2541 0.2165 0.2190 %traced_malate_422 (M3) 0.0979 0.1101 0.1012 0.1536 0.1549 0.1551 0.1250 0.0804 0.1044 0.1095 0.1022 0.1080 0.0880 0.0963 0.0872 0.0979 0.1101 0.1012 0.1360 0.1400 0.1947 0.1017 0.1002 0.1207 %traced_malate_423 (M4) 0.0221 0.0375 0.0303 0.0164 0.0359 0.0676 0.0177 0.0013 0.0162 0.0206 0.0176 0.0171 0.0298 0.0278 0.0221 0.0221 0.0375 0.0303 0.0349 0.0394 0.0554 0.0259 0.0214 0.0166 %traced_Pyruvate_174 (M0) 0.6398 0.5866 0.6261 0.5891 0.5685 0.6227 0.6233 0.6665 0.6505 0.6521 0.6886 0.6460 0.8782 0.8484 0.8492 0.8037 0.8398 0.8083 0.8049 0.7297 0.7070 0.7453 0.7538 0.7709 %traced_Pyruvate_175 (M1) 0.0391 0.0440 0.0955 0.0461 0.0425 0.0456 0.0248 0.0277 0.0306 0.0307 0.0258 0.0377 0.0887 0.0820 0.0837 0.0821 0.0943 0.0999 0.1048 0.1264 0.1293 0.1388 0.1356 0.1373 %traced_Pyruvate_176 (M2) 0.0528 0.0657 0.0259 0.0656 0.0661 0.0593 0.0491 0.0496 0.0521 0.0492 0.0470 0.0592 0.0343 0.0484 0.0371 0.0360 0.0434 0.0334 0.0209 0.0467 0.0470 0.0562 0.0558 0.0488 %traced_Pyruvate_177 (M3) 0.2710 0.3060 0.3082 0.3007 0.3244 0.2739 0.3046 0.2579 0.2689 0.2683 0.2412 0.2581 0.1711 0.1723 0.1773 0.1566 0.1551 0.1558 0.1745 0.2094 0.2383 0.1607 0.1558 0.1439 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name pubchem_id inchi_key kegg_id other_id other_id_type ri ri_type moverz_quant Alanine_total Aspartate_total Citrate_total Lactate_total Malate_total Pyruvate_total %traced_Alanine_260 (M0) 260 %traced_Alanine_261 (M1) 261 %traced_Alanine_262 (M2) 262 %traced_Alanine_263 (M3) 263 %traced_Aspartate_418 (M0) 418 %traced_Aspartate_419 (M1) 419 %traced_Aspartate_420 (M2) 420 %traced_Aspartate_421 (M3) 421 %traced_Aspartate_422 (M4) 422 %traced_Citrate_591 (M0) 591 %traced_Citrate_592 (M1) 592 %traced_Citrate_593 (M2) 593 %traced_Citrate_594 (M3) 594 %traced_Citrate_595 (M4) 595 %traced_Citrate_596 (M5) 596 %traced_Citrate_597 (M6) 597 %traced_lactate_261 (M0) 261 %traced_lactate_262 (M1) 262 %traced_lactate_263 (M2) 263 %traced_lactate_264 (M3) 264 %traced_malate_419 (M0) 419 %traced_malate_420 (M1) 420 %traced_malate_421 (M2) 421 %traced_malate_422 (M3) 422 %traced_malate_423 (M4) 423 %traced_Pyruvate_174 (M0) 174 %traced_Pyruvate_175 (M1) 175 %traced_Pyruvate_176 (M2) 176 %traced_Pyruvate_177 (M3) 177 METABOLITES_END #END