RNA from hearts or NRVMs was extracted with Trizol reagent (Invitrogen), cDNA synthesis utilized the SuperScript first-strand synthesis system (Bio-Rad), and quantity of cDNA for each transcript was measured using real-time PCR with the SYBER green supermix (Bio-Rad), and the primer sequences of PCR for α-MHC, β-MHC, Foxo1, and cyclophilin as previously described9 (link), 15 (link).
Protocol detail
Gene Expression Analysis in Cardiac Cells
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Corresponding Organization :
Other organizations : Central Texas Veterans Health Care System, North China University of Science and Technology
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Variable analysis
independent variables
- RNA source (hearts or NRVMs)
- Quantity of cDNA for each transcript (α-MHC, β-MHC, Foxo1, and cyclophilin)
- Trizol reagent (Invitrogen) for RNA extraction
- SuperScript first-strand synthesis system (Bio-Rad) for cDNA synthesis
- SYBER green supermix (Bio-Rad) for real-time PCR
- Primer sequences for α-MHC, β-MHC, Foxo1, and cyclophilin as previously described
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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