Protein Expression Profiling in Cardiomyocytes

Cytosolic and mitochondrial proteins were harvested from cardiomyocytes or heart homogenates, and protein concentration detected. The levels of specific proteins were determined by western blotting assay, as previously described [23 (link)]. The following primary antibodies were used: anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH), anti-voltage-dependent anion channel (VDAC), anti-c-jun NH2-terminal kinase (JNK), anti-extracellular signal-regulated kinase (ERK), anti-p38 MAPK (mitogen-activated protein kinase), anti-IκBα, anti-Ca2+/calmodulin-dependent protein kinase II (CaMKII), anti-p-JNK, anti-p-ERK, anti-p-p38 MAPK, anti-p-IκBα, anti-p-CaMKII, anti-Bcl-2, anti-cytochrome c (Cyt c), anti-Bax and anti-TNF-α antibody (Cell Signaling technology, Beverly, MA, USA). GAPDH or VDAC was used to normalize protein loading.