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MB Sample ID: SA055258
| Local Sample ID: | 8 |
| Subject ID: | SU000962 |
| Subject Type: | Cell culture |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Cell Strain Details: | C57BL/6 |
| Cell Primary Immortalized: | AT1 |
| Cell Counts: | ~60% confluency 6 well plate |
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Collection:
| Collection ID: | CO000956 |
| Collection Summary: | Media harvested at 14, 30, and 60 min post stretch (1Hz, 15%) |
| Collection Protocol ID: | None |
| Collection Protocol Comments: | Non-targeted metabolomics analysis. After HL-1 cells were cultured for 16 h in specialized 6well BioFlex culture plates in Claycomb medium, medium was changed to DMEM supplemented 1% penicillin-streptomycin and 10% serum. Transient knockdown of MuRF1 was carried out using recombinant Ad.shRNA MuRF1 at MOI 30. After transduction for 48 h, HL-1 cells were used for mechanical stretch at 15% strain and for different times (15, 30, and 60 min) in a computer-regulated Flexcell FX-5000 Compression System (Flexcell International Corporation, Hillsborough, NC). Non-stretch cells (0 min) were used as the control. After HL-1 cells were stretched, HL-1 cells (at 0 and 60 min) and DMEM medium (at 0, 15, 30, and 60 min) were collected for GC/MS measurement of metabolites, and samples were placed on dry ice/stored at -80C. Samples were then analyzed by GC/MS, as previously described [24]. The raw, transformed, and sorted data used for each of the three comparisons in the metabolomics analyses can be found in Supplemental Table 1. Up to 1 missing value per group was imputed using lowest value in the same group, with groups missing 2 or more excluded from the analysis. The data obtained in this study will be accessible at the NIH Common Fund’s Data Repository and Coordinating Center (supported by NIH grant, U01-DK097430) website, http://www.metabolomicsworkbench.org. |
| Sample Type: | Media |
| Collection Method: | Pipette |
| Collection Location: | Tissue culture. |
| Collection Frequency: | q 15-30 minutes |
| Collection Duration: | 1 hour |
| Volumeoramount Collected: | 10 ul |
| Storage Conditions: | -80C |
| Collection Vials: | Cryovials |
| Storage Vials: | Cryovials |
| Collection Tube Temp: | Ice |
