Anti phospholamban

Western blot analyses were carried out as previously described56 (link). The primary antibodies anti-Phospho-phospholamban (Ser16, Cell Signaling, Beverly, MA, USA), anti-phospholamban (Abcam, Cambridge, MA, USA), anti-Phospho-lamin A (Ser22, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-lamin A (Abcam) and anti-β-actin (Cell Signaling) were used.

Western blot analyses were carried out as described previously [26] (link). Briefly, left ventricular free walls were lysed with the radioimmunoprecipitation assay buffer, and the lysates were separated by SDS-PAGE. The separated proteins were then transferred to Polyvinylidene fluoride membranes. The membrane blots were first probed with a primary antibody overnight at 4°C and then secondary antibody coupled to horseradish peroxidase. The proteins were visualized with the enhanced chemiluminescent system (Pierce, Rockford, IL) and the bands densitometry was analyzed. Phosphoprotein levels were normalized to total protein levels. The primary antibodies used included anti-Phospho-phospholamban (Ser16, Cell Signaling, Beverly, MA, USA), anti-phospholamban (Abcam, Cambridge, MA, USA), anti-Phospho-lamin A (Ser22, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and anti-lamin A (Abcam).