Perihematomal brain tissues were harvested on day 3 after operation. The total proteins, nuclear and cytoplasmic proteins, and mitochondrial proteins were extracted with the RIPA lysis buffer (Millipore, Bedford, MA, USA), Nuclear and Cytoplasmic Protein Extraction Kit (Beyotime Biotechnology, Haimen, Jiangsu, China), and Mitochondrial Protein Extraction Kit (BestBio, Shanghai, China), respectively. The quantitative proteins were loaded onto 10% sodium dodecyl sulfate-polyacrylamide gel by electrophoresis (SDS-PAGE) and wet-transferred to a 0.22 or 0.45 μm PVDF membrane (Millipore, Temecula, CA, USA). At room temperature, the immunoblots were blocked by 5% nonfat milk for 1 h. Then, the immunoblots were probed with the target primary antibodies against PARP1 (1 : 5,000 dilution, catalog no. ab227244), Bcl-2 (1 : 1,000 dilution, catalog no. ab194583), NLRP3 (1 : 1,000 dilution, catalog no. ab4207), caspase-1 (1 : 1,000 dilution, catalog no. ab1872), IL-18 (1 : 400 dilution, catalog no. ab71495), NF-κB (1 : 1,000 dilution, catalog no. ab19870), Nrf2 (1 : 1,000 dilution, catalog no. ab137550), OPTN (1 : 300 dilution, catalog no. ab23666), Beclin1 (1 : 2,000 dilution, catalog no. ab207612), Parkin (1 : 500 dilution, catalog no. ab77924), CoxIV (1 : 2,000 dilution, catalog no. Ab33985), and β-actin (1 : 5,000 dilution, catalog no. ab8227, all from Abcam, Cambridge, UK); Bax (1 : 1,000 dilution, catalog no. 2772), ASC (1 : 1,000 dilution, catalog no. 67824), IL-1β (1 : 1,000 dilution, catalog no. 52718), LC3 (1 : 1,000 dilution, catalog no. 4108), PINK1 (1 : 1,000 dilution, catalog no. 6946), and Histone H3 (1 : 1,000 dilution, catalog no. 4499, all from Cell Signaling Technology, Beverly, MA, USA) at 4°C overnight. After incubation with the corresponding secondary antibodies (1 : 5,000 dilution, Hua An, Hangzhou, Zhejiang, China) for another 1 h at room temperature, the ECL chemiluminescence (CL) reagent (Pierce, Rockford, IL, USA) was employed to detect the protein signals. The final results were quantified with the Quantity One software (Bio-Rad, Hercules, CA, USA).

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