Quantitative PCR analysis
Quantitative PCR (qPCR) was performed as previously described (Jin et al., 2020). Briefly, total RNA was extracted using TRizol (Life Technologies, Frederick, MD, USA), followed by ethanol extraction. qPCR was conducted in triplicate using SYBR Green PCR reagent (Bio-Rad, Hercules, CA, USA) on the CFX96 Real-Time PCR system (Bio-Rad). PCR conditions included a denaturation step of 95°C for 5 min, followed by 40 cycles consisting for 95°C for 15 sec, 60°C for 30 sec, and 72°C for 30 sec. The primer sequences are described in Supplementary Table 2. The relative expression of each target gene was calculated by the 2-ΔΔCt method after normalization to β-Actin.