Western Blot Analysis of Cardiac Proteins

Proteins were isolated from the ventricular homogenate or cells with lysis buffer (Beyotime Institute of Biotechnology, Shanghai, China) containing phenylmethylsulfonyl fluoride (PMSF) (Sigma-Aldrich) and subjected to Western blot, as described previously [24 (link)]. The antibodies used in this study were glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and collagen type III (Col3al) (Proteintech, Rosemont, IL, USA), matrix metalloproteinase 2 (MMP2) and MMP9 (Abcam, Cambridge, MA, USA), α-SMA (Bioss, Beijing, China), cellular communication network factor 2 (CCN2) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and TNF-α, p-AKT (Ser473), AKT, p-ERK (Thr202/Tyr204), ERK, p-STAT3 (Tyr705), STAT3, and Caspase-3 (Cell Signaling Technology, Danvers, MA, USA).