RT-PCR splicing analysis

RTn was performed using 1 μg total cellular RNA, Superscript II (ThermoFisher), and oligo dT₁₅ plus random hexamer primers, per the manufacturer's protocol. PCR was then performed using 5′ FAM-labeled primers flanking mSerca1 exon 22 and mClcn1 exon 7A in quadriceps cDNA (18,31), or mTmem63b exon 5 and mCacna1s exon 29 in heart cDNA (32) (Table ​(Table1).1). The PCR products were separated by agarose gel electrophoresis and detected with a laser fluorimager (Typhoon, GE Healthcare). The products were quantified using ImageQuant software (GE Healthcare).