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MB Sample ID: SA204348
| Local Sample ID: | HSC_Cultured_6h_2 |
| Subject ID: | SU002210 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002210 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| HSC_Cultured_6h_2 | SA204348 | FL025085 | Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Source_Name[gating] |
| HSC_Cultured_6h_2 | SA204348 | FL025085 | Ex vivo culture 2 weeks, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Treatment |
| HSC_Cultured_6h_2 | SA204348 | FL025085 | 1b | Batch |
| HSC_Cultured_6h_2 | SA204348 | FL025085 | [13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig2.D). | Note |
Collection:
| Collection ID: | CO002203 |
| Collection Summary: | 5×10^5 indicated cells were incubated with 100 µM stable [13C,15N] amino acids (MSK-A2-US-1.2, Cambridge Isotope Laboratories) for indicated time and centrifuged at 500 g for 5 min at 4 °C. The pelleted cells were extracted in 500 µl ice-cold Acetonitrile: Isopropyl Alcohol: water (3:3:2 v/v/v) and aliquoted as three technical replicates. The extracts were vortexed for 5 min at 4 °C and centrifuged at 14,000 g for 2 min at 4 °C. The supernatants were dried by vacuum spin for subsequent derivatization and stored at -20 ℃. |
| Sample Type: | Bone marrow |
Treatment:
| Treatment ID: | TR002222 |
| Treatment Summary: | Germ-free C57BL/6J mice were intraperitoneally injected with 10 mg/kg 5FU (F6627-5G, Sigma-Aldrich) for 14 days, or fed with vehicle or NR (400 mg/kg per day) (1341-23-7, ziyi-reagent) for consecutive 8 weeks (long-term) or 1 week (short-term). Hematopoietic cells were stained and sorted from treated mice. |
Sample Preparation:
| Sampleprep ID: | SP002216 |
| Sampleprep Summary: | To get the corresponding derivatives, the dried aliquots were incubated with 20 µL 2% (w/v) methoxyamine hydrochloride (226904, Sigma-Aldrich) in pyridine for 60 min at 37 °C, and silylated by 30 µL of N-Methyl-N-(tert-butyldimethylsilyl) trifluoroacetamide with 1% tert-Butyldimethylchlorosilane (TBDMS, 18162-48-6, Regis Technologies) for 30 min at 45 °C. The corresponding derivatives were analyzed by GC-MS using the Trace 1310 gas chromatograph (Thermo Fisher) with the DB-35ms column (Agilent Technologies) connected to the Q ExactiveTM GC OrbitrapTM GC-MS/MS system (Thermo Fisher). |
Combined analysis:
| Analysis ID | AN003478 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | TRACE 1310 |
| Column | DB-35ms |
| MS Type | EI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE |
| Units | Peak Area |
Chromatography:
| Chromatography ID: | CH002567 |
| Instrument Name: | TRACE 1310 |
| Column Name: | DB-35ms |
| Chromatography Type: | GC |
MS:
| MS ID: | MS003239 |
| Analysis ID: | AN003478 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | EI |
| MS Comments: | GC-MS data was acquired on Q-Exactive Orbitrap mass spectrometer (Thermo Fisher) coupled with Gas Chromatograph system TRACE 1310 (Thermo Fisher). Chromatographic separation was performed on a DB-35ms column module (30 m length x 0.25 mm internal diameter, Agilent Technologies). The column temperature was programmed with an initial temperature of 50 °C for 2 min, then ramped at 10°C/min to 325 °C, and maintained for 5 min. The mass range was set as 50-600 m/z, and the resolution was 60,000. The ion source temperature was 300°C with the transfer line temperature of 250°C, and the electron energy was 70 eV with EI source. |
| Ion Mode: | POSITIVE |
