Anti lc3 2 lc3 1

Each cell group was collected and lysed in RIPA buffer (Biosharp, HeFei, China) to complete protein extraction. The collected protein samples were added to a sodium dodecyl sulfate polyacrylamide gel (Activagen Bio, Hefei, China) at a ratio of 1:4 to separate the proteins. Then, the gels were electrotransferred to polyvinylidene difluoride membranes (Millipore, USA). The membranes were blocked using 5% skim milk powder. Then, the following primary antibodies were added overnight at 4 °C: anti-β-Actin (Zs-BIO, USA, 1:1,000); anti-Pink1 (Affinity, USA, 1:1,000); anti-Parkin (Affinity, USA, 1:1,000); anti-Rab7 (ZENBIO, ChengDu, China, 1:1,000); anti-NLRP3 (Affinity, USA, 1:1,000); anti-Caspase-1 (Affinity, USA, 1:1,000); anti-Podxl (ABclonal, WuHan, China, 1:1,000); anti-Beclin1 (Affinity, USA, 1:500); anti-LC3 II/LC3 I (Affinity, USA, 1:500); and anti-P62 (CST, USA, 1:1000). The membranes were washed with PBS buffer and incubated with the secondary antibody for 2 h at room temperature, after which the membrane signal was observed via an enhanced chemiluminescence kit (Thermo, USA). Film strips were analyzed using ImageJ software 1.8.0.