Immunoblotting was performed as described previously [4 (link)]. Protein concentrations in mitochondria and homogenates were determined using a Bradford assay kit (Bio-Rad, Hercules, CA, USA). Equal amounts of mitochondrial and homogenate proteins from the different experimental groups were resolved on 10% SDS-PAGE and transferred onto Amersham Hybond ECL nitrocellulose membranes (GE Healthcare Bio-Sciences Corp., Pittsburgh, PA). Membranes were blocked with 5% BSA in TBS. The antibodies used in this study were against total forms of PKCɛ (1:1000), PKCδ (1:200), AT1-R (1:200), AT2-R (1:200) (Santa Cruz Biotechnology, Santa Cruz, CA), Akt (1:1000), ERK1/2 (1:1000), JNK (1:1000), p38 (1:1000), PKG-1 (1:1000), VDAC (1:1000) (Cell Signaling Technology, Danvers, MA), and GAPDH (1:10000) (Sigma-Aldrich, MO, USA). The signals were visualized by Odyssey CLx Quantitative Fluorescent Imaging System with the secondary infrared antibodies IRDye 800CW goat anti-rabbit (1:25000) and 800CW donkey anti-goat (1:25000). Results were analyzed with Image Studio Lite Software (LI-COR Biotechnology, Lincoln, NE).