qPCR

Total RNA (day 14 posttransfection) was extracted using Quick-RNA MiniPrep Kit according to the manufacturer’s instructions (Zymo Research). Total RNA (50–100 ng) was converted to cDNA using a high-capacity cDNA reverse transcription kit (Applied Biosystems). cDNA was used in a standard qPCR reaction involving FAM-conjugated gene-specific primers (Thermo Fisher) and TaqMan Gene Expression Master Mix (Thermo Fisher). Primers were acquired from Thermo Fisher and the assay ID numbers are Actn2 Mm00473657_m1, Gapdh Mm99999915_m1, Myh6 Mm00440359_m1, and Tnni3 Mm00437164_m1.