2.4. Effects of silencing GSDMD by siRNA on pyroptosis in MG cells

To investigate the effects of GSDMD on pyroptosis and inflammatory factor release in MG cells, siRNA (GenePharm Co., Ltd.) was used to silence GSDMD expression. The verified siRNA sequence with relatively good silencing efficacy was as follows: CTCCATGAATGTGTGTATACT. MG cells were seeded into 24‐well plates at confluency of 30%. The total medium before transfection was 0.45 ml. A 50 pmol (0.67 μg) of siRNA was added to Opti‐MEM (Sigma) at a final volume of 25 μl. A 1 μl of Entranster‐R4000 was added to 24 μl of Opti‐MEM, followed by incubation at room temperature for 5 min. The diluted siRNA and the diluted Entranster‐R4000 dilution were mixed for incubation at room temperature for 15 min. A 50 μl of the transfection solution was added to 0.45 ml of complete medium to mix well. After transfection for 6 hr, the MG cell state was good. MG cells were incubated for 48 hr, followed by detection of the expression level of GSDMD mRNA.