#METABOLOMICS WORKBENCH araskind_20160412_125342 DATATRACK_ID:580 STUDY_ID:ST000370 ANALYSIS_ID:AN000604 PROJECT_ID:PR000129 VERSION 1 CREATED_ON April 18, 2016, 4:03 pm #PROJECT PR:PROJECT_TITLE Metabolic effects of Noxa PR:PROJECT_SUMMARY labeled glutamine metabolomics on Noxa expressing cells PR:INSTITUTE Masonic Cancer Center, University of Minnesota PR:DEPARTMENT Masonic Cancer Center PR:LABORATORY Kelekar Lab PR:LAST_NAME Kelekar PR:FIRST_NAME Ameeta PR:ADDRESS 420 Delaware Street SE, Minneapolis, MN 55455 PR:EMAIL ameeta@umn.edu PR:PHONE 612-626-2358 #STUDY ST:STUDY_TITLE Primary T Cell Noxa Knockdown (Donor 8) III ST:STUDY_TYPE Regular ST:STUDY_SUMMARY Labeling of cells was carried out in triplicate with each sample containing ST:STUDY_SUMMARY 35e6. Cells were deprived of glucose or glutamine for 1 h then resuspended in ST:STUDY_SUMMARY 10mM [13C1,2] D-glucose labeled or 4 mM [13C5]-U-glutamine labeled medium, ST:STUDY_SUMMARY respectively. At this time cells were also activated with 5 ug/ml anti-CD3 and ST:STUDY_SUMMARY anti-CD28. Cells were labeled (and activated) for 16 h when samples were spun ST:STUDY_SUMMARY down, and cells were resuspended in 200 ul ice cold methanol. For this part of ST:STUDY_SUMMARY the experiment, the spent medium was collected for analysis. All samples were ST:STUDY_SUMMARY immediately stored in -80oC. ST:INSTITUTE University of Michigan ST:DEPARTMENT Biomedical Research ST:LABORATORY Core Facilities Metabolomics core ST:LAST_NAME Kachman ST:FIRST_NAME Maureen ST:ADDRESS 6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714 ST:EMAIL mkachman@med.umich.edu ST:PHONE (734) 232-8175 ST:NUM_GROUPS 3 ST:TOTAL_SUBJECTS 16 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS SU0012050 S00017964 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0012051 S00017965 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0012052 S00017966 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0012053 S00017967 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0012054 S00017968 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0012055 S00017969 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0012056 S00017970 Glucose label:no | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0012057 S00017971 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0012058 S00017972 Glucose label:yes | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0012059 S00017973 Glucose label:yes | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0012060 S00017974 Glucose label:yes | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0012061 S00017975 Glucose label:yes | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0012062 S00017976 Glucose label:yes | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0012063 S00017977 Glucose label:yes | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0012056 S00017978 Glucose label:no | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0012064 S00017979 Glucose label:yes | Glutamine label:no #COLLECTION CO:COLLECTION_SUMMARY None CO:SAMPLE_TYPE Culture Media #TREATMENT TR:TREATMENT_SUMMARY None #SAMPLEPREP SP:SAMPLEPREP_SUMMARY None SP:SAMPLEPREP_PROTOCOL_FILENAME Fluxomics analysis protocol-2015-03-11.docx #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 7890A CH:COLUMN_NAME Agilent DB5-MS (30m × 0.25mm, 0.25um) CH:METHODS_FILENAME DQM0208 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent 5975C MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Area MS_METABOLITE_DATA_START Samples S00017964 S00017965 S00017966 S00017967 S00017968 S00017969 S00017970 S00017971 S00017972 S00017973 S00017974 S00017975 S00017976 S00017977 S00017978 S00017979 Factors Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:no Glucose label:no | Glutamine label:yes Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:no | Glutamine label:no Glucose label:yes | Glutamine label:no Aspartate _1234 9315.2 39815.0 16855.3 105275.1 108065.6 101531.6 95095.1 109268.8 80174.0 95234.6 105221.5 58153.4 40300.6 88247.7 98286.0 95446.6 Aspartate _1234_13C_1 163.7 414.8 741.0 129.6 613.4 1322.4 1179.7 6650.4 2777.2 534.5 1203.7 159.1 Aspartate _1234_13C_2 598.0 97.3 604.7 96.0 510.7 254.7 609.1 2.0 239.8 340.5 1195.3 Aspartate _1234_13C_3 648.6 876.1 444.4 159.4 176.6 882.1 149.2 479.4 41.6 Aspartate _1234_13C_4 2967.2 2056.7 2788.6 131.6 83.9 43.0 22.3 147.8 86.6 Lactate_123 6361118.7 4135958.7 5921024.1 161314.4 206333.0 185722.2 14659.9 18532.9 650301.8 519693.9 500979.3 710505.9 813099.8 468961.2 16255.5 10116.7 Lactate_123_13C_1 9254.1 17118.2 27882.9 441.6 347.3 423.4 45.0 30369.6 27200.4 26542.4 31965.7 44127.8 22342.2 204.7 24.1 Lactate_123_13C_2 9366.8 2430.4 15325.5 1054.8 818.1 1037.7 558072.0 457561.6 446911.1 608078.6 680792.8 411714.2 2421.2 568.8 Lactate_123_13C_3 53806.2 27858.7 37259.5 421.6 342.0 431.2 284.7 11642.4 8484.6 4941.8 9377.0 7711.2 462.5 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID Lactate_123 612 Lactate_123_13C_1 612 Lactate_123_13C_2 612 Lactate_123_13C_3 612 Aspartate _1234 424 Aspartate _1234_13C_1 424 Aspartate _1234_13C_2 424 Aspartate _1234_13C_3 424 Aspartate _1234_13C_4 424 METABOLITES_END #END