RNA extraction and real-time qPCR
Total RNA was extracted from ventricular apical myocardium using TRIzol
(15596026; Life technologies). cDNA was synthesized from 1ug of RNA
using a Reverse Transcription Reagent kit according to the
manufacturer’s instruction (FSQ107; Toyobo). The obtained cDNAs were
amplified by semiquantitative RT-PCR using SYBR Green Master Mix
(Q111-02; Vazyme). The primer sequences used are listed here
((Supplementary Table S1). The relative gene expressions were normalized
to the of GAPDH and comparisons were done with the ddCt method as
described (Gold JI et al., 2013).