Anti pkcε antibody

Left ventricles were dissected out from mice 5 days after SNI surgery and homogenized in buffer A (in mM: 5 Tris, 4 EGTA, 2 EDTA, 5 DTT, 1 PMSF) and containing EDTA-free protease inhibitor and vortex every 5 min and stood on ice for 20 min. The homogenate was centrifuged at 100,000 × g for 30 min at 4 °C and the supernatant was collected as the cytosolic fraction. The remaining pellet was resuspended and incubated with buffer B (buffer A containing 1% Triton X-100) on ice for 20 min and then centrifuged at 100,000 × g for 30 min at 4 °C. This supernatant was collected as the membrane-associated fraction. Protein concentration was determined by Bradford dye-binding method (Bio-rad). Anti-PKCε antibody (1: 1000, Cell Signaling), anti-cleaved caspase 3 (1:1000, Cell Signaling) antibody were used to detect PKCε and cleaved caspase 3, respectively. Anti-GAPDH (1:10,000, Sigma) and anti-α1-sodium/potassium ATPase (1:1000, Santa Cruz) antibodies were used for housekeeping proteins. Uncropped images of the blots are presented in Supplementary Fig. 7.