Neonatal rats were purchased from Charles River Laboratories, and NRVMs were isolated using previously published techniques (45 (link)). NRVMs were serum starved for 6 hours before 18-hour treatment, unless otherwise specified. The following reagents were used: activin A (0 to 100 ng/ml: 338 AC, R&D Systems), GDF11 (100 ng/ml: 120–11, PeproTech), GDF8 (100 ng/ml: 788G8, R&D Systems), CDD866 (0 to 100 μg/ml; Novartis), MG132 (10 μM; Sigma-Aldrich), and A01 (10 μM; Sigma-Aldrich). Proteasome activity was measured using the 20S Proteasome Activity Assay (MilliporeSigma).
Protocol detail
Neonatal Rat Ventricular Myocyte Isolation and Treatment
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Variable analysis
independent variables
- Activin A (0 to 100 ng/ml)
- GDF11 (100 ng/ml)
- GDF8 (100 ng/ml)
- CDD866 (0 to 100 μg/ml)
- MG132 (10 μM)
- A01 (10 μM)
- Proteasome activity
- Neonatal rats from Charles River Laboratories
- NRVMs isolated using previously published techniques
- NRVMs serum starved for 6 hours before 18-hour treatment, unless otherwise specified
- Not explicitly mentioned
- Not explicitly mentioned
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