qPCR Quantification of Target Genes

Analyses of q-PCR were performed as previously described [20 (link)]. Total RNA was extracted using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol. HiScript Q RT SuperMix for q-PCR kit (Vazyme, Nanjing, China) was used for reverse transcription polymerase chain reactions, and then q-PCR assays were conducted with SYBR Green I mix (Takara, Dalian, China) on an ABI ViiA 7 Q-PCR System (Applied Biosystems, Waltham, MA). In all cases, mRNA levels were normalized to the expression of GAPDH, which served as an endogenous control. The relative expression of target genes was calculated by the 2^-△△Ct method [22 (link)]. The primer sets used in this study are listed in Additional file 1: Table S2.