#METABOLOMICS WORKBENCH Advanilab_metabolomics_20230107_131701 DATATRACK_ID:3684 STUDY_ID:ST002437 ANALYSIS_ID:AN003971 VERSION 1 CREATED_ON 02-08-2024 #PROJECT PR:PROJECT_TITLE Metabolomic analysis in wildtype and Isg15 knockout mice after sham or PR:PROJECT_TITLE transverse aortic constriction surgery PR:PROJECT_TYPE Untargeted metabolomic analysis PR:PROJECT_SUMMARY Left ventricular tissue was isolated from wildtype and Isg15 knockout mice 8 PR:PROJECT_SUMMARY weeks after sham or transverse aortic constriction surgery and the metabolomic PR:PROJECT_SUMMARY differences across the samples were compared. PR:INSTITUTE St. Michael's Hospital PR:DEPARTMENT Keenan Research Centre for Biomedical Science PR:LABORATORY Advani Lab PR:LAST_NAME Advani PR:FIRST_NAME Andrew PR:ADDRESS 6-151, 61 Queen St. East, Toronto, Ontario. M5C 2T2 PR:EMAIL andrew.advani@unityhealth.to PR:PHONE 4163604000 PR:DOI http://dx.doi.org/10.21228/M8WD9H #STUDY ST:STUDY_TITLE Metabolomic analysis in wildtype and Isg15 knockout mice after sham or ST:STUDY_TITLE transverse aortic constriction surgery ST:STUDY_SUMMARY Left ventricular tissues were isolated from wildtype and Isg15 knockout mice 8 ST:STUDY_SUMMARY weeks after sham or transverse aortic constriction and the metabolomic ST:STUDY_SUMMARY differences were compared. ST:INSTITUTE St. Michael's Hospital ST:LAST_NAME Advani ST:FIRST_NAME Andrew ST:ADDRESS 6-151, 61 Queen Street East, Toronto, Ontario. M5C 2t2 ST:EMAIL andrew.advani@unityhealth.to ST:PHONE 4163604000 ST:SUBMIT_DATE 2023-01-07 #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 - Sample 10 Genotype:ISG15 KO | Surgical condition:Sham RAW_FILE_NAME=VQE001982_EP_Advani_+_10.raw; RAW_FILE_NAME=VQE002012_EP_Advani_-_10.raw SUBJECT_SAMPLE_FACTORS - Sample 6 Genotype:ISG15 KO | Surgical condition:Sham RAW_FILE_NAME=VQE001978_EP_Advani_+_6.raw; RAW_FILE_NAME=VQE002008_EP_Advani_-_6.raw SUBJECT_SAMPLE_FACTORS - Sample 7 Genotype:ISG15 KO | Surgical condition:Sham RAW_FILE_NAME=VQE001979_EP_Advani_+_7.raw; RAW_FILE_NAME=VQE002009_EP_Advani_-_7.raw SUBJECT_SAMPLE_FACTORS - Sample 8 Genotype:ISG15 KO | Surgical condition:Sham RAW_FILE_NAME=VQE001980_EP_Advani_+_8.raw; RAW_FILE_NAME=VQE002010_EP_Advani_-_8.raw SUBJECT_SAMPLE_FACTORS - Sample 16 Genotype:ISG15 KO | Surgical condition:TAC RAW_FILE_NAME=VQE001988_EP_Advani_+_16.raw; RAW_FILE_NAME=VQE002018_EP_Advani_-_16.raw SUBJECT_SAMPLE_FACTORS - Sample 17 Genotype:ISG15 KO | Surgical condition:TAC RAW_FILE_NAME=VQE001989_EP_Advani_+_17.raw; RAW_FILE_NAME=VQE002019_EP_Advani_-_17.raw SUBJECT_SAMPLE_FACTORS - Sample 18 Genotype:ISG15 KO | Surgical condition:TAC RAW_FILE_NAME=VQE001990_EP_Advani_+_18.raw; RAW_FILE_NAME=VQE002020_EP_Advani_-_18.raw SUBJECT_SAMPLE_FACTORS - Sample 19 Genotype:ISG15 KO | Surgical condition:TAC RAW_FILE_NAME=VQE001991_EP_Advani_+_19.raw; RAW_FILE_NAME=VQE002021_EP_Advani_-_19.raw SUBJECT_SAMPLE_FACTORS - Sample 2 Genotype:Wild type | Surgical condition:Sham RAW_FILE_NAME=VQE001974_EP_Advani_+_2.raw; RAW_FILE_NAME=VQE002004_EP_Advani_-_2.raw SUBJECT_SAMPLE_FACTORS - Sample 3 Genotype:Wild type | Surgical condition:Sham RAW_FILE_NAME=VQE001975_EP_Advani_+_3.raw; RAW_FILE_NAME=VQE002005_EP_Advani_-_3.raw SUBJECT_SAMPLE_FACTORS - Sample 4 Genotype:Wild type | Surgical condition:Sham RAW_FILE_NAME=VQE001976_EP_Advani_+_4.raw; RAW_FILE_NAME=VQE002006_EP_Advani_-_4.raw SUBJECT_SAMPLE_FACTORS - Sample 11 Genotype:Wild type | Surgical condition:TAC RAW_FILE_NAME=VQE001983_EP_Advani_+_11.raw; RAW_FILE_NAME=VQE002013_EP_Advani_-_11.raw SUBJECT_SAMPLE_FACTORS - Sample 12 Genotype:Wild type | Surgical condition:TAC RAW_FILE_NAME=VQE001984_EP_Advani_+_12.raw; RAW_FILE_NAME=VQE002014_EP_Advani_-_12.raw SUBJECT_SAMPLE_FACTORS - Sample 13 Genotype:Wild type | Surgical condition:TAC RAW_FILE_NAME=VQE001985_EP_Advani_+_13.raw; RAW_FILE_NAME=VQE002015_EP_Advani_-_13.raw SUBJECT_SAMPLE_FACTORS - Sample 14 Genotype:Wild type | Surgical condition:TAC RAW_FILE_NAME=VQE001986_EP_Advani_+_14.raw; RAW_FILE_NAME=VQE002016_EP_Advani_-_14.raw #COLLECTION CO:COLLECTION_SUMMARY Mouse tissue frozen samples were reconstituted in 500 uL 80% MeCN containing 1 CO:COLLECTION_SUMMARY uM caffeine-d9 internal standard. The samples were homogenized, vortexed and CO:COLLECTION_SUMMARY centrifuged 21000 x g for two minutes at room temperature (23°C). CO:SAMPLE_TYPE Cardiac tissue #TREATMENT TR:TREATMENT_SUMMARY Wildtype and ISG15 knockout mice underwent sham or transverse aortic TR:TREATMENT_SUMMARY constriction surgery. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY 200 uL of each sample supernatants were diluted with 200 uL H2O and 100 uL used SP:SAMPLEPREP_SUMMARY for the LC-MS analysis. LC-MS analyses were performed using Vanquish UPLC with SP:SAMPLEPREP_SUMMARY 0-100% 10 minutes gradient (0.1% FA - 0.1%FA/MeCN) at 200 uL/min flow rate on SP:SAMPLEPREP_SUMMARY Phenomenex Luna Omega 3 um Polar C18 100A 50x2.1 mm (RP-LC-MS). 10 uL injection SP:SAMPLEPREP_SUMMARY volumes were used for each run. Mass spectra were acquired using Thermo Orbitrap SP:SAMPLEPREP_SUMMARY QE+ spectrometer in positive and negative modes in separate runs. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY LC-MS analyses were performed using Vanquish UPLC with 0-100% 10 minutes CH:CHROMATOGRAPHY_SUMMARY gradient (0.1% FA - 0.1%FA/MeCN) at 200 uL/min flow rate on Phenomenex Luna CH:CHROMATOGRAPHY_SUMMARY Omega 3 um Polar C18 100A 50x2.1 mm (RP-LC-MS). 10 uL injection volumes were CH:CHROMATOGRAPHY_SUMMARY used for each run. CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Phenomenex Luna Omega Polar C18(50 x 2.1mm, 3um. 100A) CH:COLUMN_TEMPERATURE 23 CH:FLOW_GRADIENT 0-100% 10 minutes gradient CH:FLOW_RATE 200 uL/min CH:SOLVENT_A 100% water; 0.1% formic acid CH:SOLVENT_B 100% acetonitrile; 0.1% formic acid CH:CHROMATOGRAPHY_TYPE Reversed phase #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Plus Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:MS_COMMENTS Mass spectra were acquired using Thermo Orbitrap QE+ spectrometer in positive MS:MS_COMMENTS and negative modes in separate runs. Data for each LC/acquisition type were MS:MS_COMMENTS analyzed using Compound Discoverer 3.3 (Thermo). MS:ION_MODE NEGATIVE MS:MS_RESULTS_FILE ST002437_AN003971_Results.txt UNITS:Peak Area Has m/z:Yes Has RT:Yes RT units:Minutes #END