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MB Sample ID: SA055256
Local Sample ID: | 19 |
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 |