Real-time qRT-PCR analysis of StUSP genes

BIOER’s LineGene 9620 real-time fluorescence quantitative PCR system was used to select 10 genes for qRT-PCR analysis, and β-action was used as the reference gene (Table S11). These genes included StUSP2, StUSP6, StUSP13, StUSP14, StUSP15, StUSP21, StUSP24, StUSP9, StUSP33, and StUSP41; the primer sequences were designed using prime-blast [89 (link)]. Total RNA was extracted from each treated sample using the Total RNA Rapid Extraction Kit (ER501-01, TransGen Biotech, Beijing, China) and reverse-transcribed to cDNA using TransScript^® One-Step gDNA Removal and cDNA Synthesis SuperMix (AT311, TransGen Biotech, Beijing, China). qRT-PCR was performed using the ChamQ Universal SYBR qPCR Master Mix (Q711, Vazyme, Nanjing, China) kit. Three biological replicates were calculated using the 2^−∆∆Ct method [90 (link)].