RNA isolation and real-time polymerase chain reaction (PCR) were performed as described previously (Zhang et al., 2019b (link)). Total RNA was prepared using the Isolate II RNA Mini Kit (Bioline USA, Inc.), and cDNA was then synthesized with the iScript cDNA Synthesis Kit (Bio-Rad). Subsequently, quantification of gene expression was performed in duplicate using iQ SYBR Green Supermix (Bio-Rad) with detection on a MyiQ Real-Time PCR System (Bio-Rad). The reaction cycles used were 95°C for 5 minutes, 40 cycles at 95°C for 15 seconds, and 58°C for 1 minute, and this was followed by melt curve analysis. Relative gene expression quantification was based on the comparative threshold cycle method (∆∆Ct) with normalization of the raw data to the housekeeping gene (18S ribosomal RNA).