The qRT-PCR analysis used 10 upper bodies of the same genotype (Figure 3A), and the semi-qRT-PCR analysis used 15–20 hearts of the same genotype (Figure 4B,F,H). Total RNA was extracted using TRIzol, and cDNA was subsequently synthesized using a Transcriptor First-Strand cDNA Synthesis Kit (Roche, Basel, Switzerland). qRT-PCR was conducted using a LightCycler 480 system (Roche, Basel, Switzerland). For semi-qRT-PCR, genes of interest were amplified using a PrimeSTAR GXL kit. cDNA levels were normalized to actb2/β-actin2 levels in both the qRT-PCR and semi-qRT-PCR experiments. The primers used for semi-qRT-PCR and qRT-PCR analysis are listed in Supplementary file 1.