Beta actin actb

Western Blotting was performed as previously described [22] (link). Briefly, the protein samples were mixed with Laemmli sample buffer (1×) and boiled for 5 min. Proteins were subjected to 10% SDS-PAGE and electroblotted onto BioRad, 0.22 µM nitrocellulose membrane (BioRad Laboratories, USA), followed by blocking for 1 h at room temperature using Tris-buffered saline with 0.2% Tween 20 (TBS-T) containing 3% BSA. The membrane was incubated with primary antibodies at 4°C overnight and subsequently incubated with secondary antibodies conjugated with horseradish peroxidase (Sigma Aldrich, USA) for 2 h at RT and washed again with TBS-T. Antibody-reactive proteins were detected by enhanced chemiluminescence using Pierce ECL Plus western blotting detection reagents (Pierce, USA). Primary antibodies for GATA4 (1∶150; Santa Cruz Biotechnology, USA), Connexin43 (CX43; 1∶500; Abcam, USA), Alpha sarcomeric actin (ACTA1; 1∶500; Sigma Aldrich, USA) and Beta Actin (ACTB; 1∶1000; Sigma Aldrich, USA) were used along with goat anti-rabbit and anti-mouse horseradish peroxidase conjugated secondary antibodies (1∶10,000; Sigma Aldrich, USA).