Quantitative Real-Time PCR Analysis

After treated with DMSO or BD for 48 h, the cells were collected. Total RNA was extracted from cells using TRIzol^™ reagent (Invitrogen) according to the manufacturer’s instructions. quantity real-time PCR (qRT-PCR) was conducted as previous report (Dong et al., 2020 (link)). Briefly, the total RNA was reversely transcribed to cDNA by using the M-MLV reverse transcriptase (Promega). SYBR PreMix Ex Taq II (Vazyme Biotech, Nanjing, China) was used to carry out qRT-PCR in 20 μL reaction system. RT-qPCR reaction was carried out by using a LightCycle96 real-time PCR system (Roche). The relative mRNA expression level was calculated by 2^−ΔΔCT method. The primers used in this study are listed in Table 1.