Gabra1

Total protein in cells was collected, and the concentration of protein wasquantified with Pierce BCA Protein Assay Kit. After segregated by 10% sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) gel, extracted proteins were transferred to polyvinylidene difluoride (PVDF) membranes and blocked in Tris-buffered saline-Tween with 5% skim milk at room temperature for 60 min. Western blot analysis was performed according to standard procedures. Primary antibodies were GABRA1 (1:1000, proteintech, USA), β-actin (1:5000, proteintech, USA), and corresponding secondary antibodies were anti-mouse and anti-rabbit(proteintech, USA). X-ray film was pressed in dark. Finally, the proteins were detected by enhanced chemiluminescence (ECL), followed by expose about 60 s for scanning and measuring.

Antibodies were purchased from the following suppliers: against Catalase (#14097), phospho-DRP1 (#3455), GAPDH (#2118), CORO1A (#92904), and LC3B (#2775) from Cell Signaling Technology, Danvers MA, USA; against CD81 (#sc-23962), CD63 (#sc-5275), and Flotillin-1 (#sc-25506) from Santa Cruz Biotechnology, Dallas, TX, USA; against β-actin (#A5441) from Sigma-Aldrich, St. Louis, MO, USA; against GFAP (#610565) from BD Transduction Laboratory, San Jose, CA, USA; against von Willebrand factor (VWF) (#ab6994), PECAM-1 (#ab28364) from Abcam, Cambridge, MA, USA; anti OMG (#12701-1-AP) and GABRA1 (#12410-1-AP) from Proteintech Group, Inc., Rosemont, IL, USA; and mitochondrial complex III subunit core 1 from Invitrogen Corporation, Fredrick, MD, USA.