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MB Sample ID: SA055252
| Local Sample ID: | 30 |
| 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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Treatment:
| Treatment ID: | TR000976 |
| Treatment Summary: | Cell culture, adenovirus transduction, and MuRF1 knock-down. The cardiomyocyte-derived HL-1 cell line was maintained as previously described [11-13]. Briefly, cells were split and cultured to ~90% confluency in 6-well BioFlex culture plates (Flexcell International Corporation, Hillsborough, NC) coated with 0.02% gelatin (wt/vol) and 0.5% fibronectin (vol/vol) for ∼30 min and grown in complete Claycomb media containing 10% serum (Cat. #51800C, Sigma-Aldrich; St. Louis, MO), as previously described [13]. Media was changed to serum-free Dulbecco’s modified Eagle’s medium (DMEM) supplemented 1% penicillin-streptomycin. HL-1 cells were transduced with 30 MOI adenovirus expressing shRNA MuRF1 (Ad.shRNA MuRF1) or shRNA scramble (Ad.shRNA Scr) control (custom made by Vector Biolabs, Philadelphia, PA) to transiently knock down MuRF1, as previously described [13]. |
| Treatment: | Ad.shRNA MuRF1 (or Ad.shRNA Scramble) treatment |
| Treatment Compound: | Adenovirus |
| Treatment Route: | Media |
| Treatment Dose: | 30 MOI |
| Treatment Dosevolume: | <10 ul |
| Treatment Doseduration: | 48 hours |
| Treatment Vehicle: | DMEM |
| Cell Media: | DMEM / No serum |
| Cell Envir Cond: | Stretch |
| Cell Pct Confluence: | ~60% |
| Cell Media Lastchanged: | At start of stretch period. |
