Immunoblot Analysis of H9c2 Cardiomyocyte Proteins

H9c2 cardiomyocyte extract (5 × 10⁵ cells) was subjected to immunoblot analysis as previously described [46 (link),64 (link)], using an anti-Cd38 polyclonal antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) raised against a peptide fragment of mouse Cd38 (residues 279–301 in [46 (link)]), anti-Ryr2 polyclonal antibody (PeproTech, Canbury, NJ, USA) [67 (link)], anti-Fkbp12.6 monoclonal antibody (Santa Cruz Biotechnology) raised against the amino acids 38–108 of human/rat/mouse FKBP12.6, anti-Pten monoclonal antibody raised against the full-length human PTEN protein (Abcam, Cambridge, UK), and anti-β-actin monoclonal antibody (Sigma, St. Louis, MO, USA) raised against Ac-Asp-Asp-Asp-Ile-Ala-Ala-Leu-Val-Ile-Asp-Asn-Gly-Ser-Gly-Lys. A SNAP id^® 2.0 Protein Detection System (Merck Millipore, Burlington, MA, USA) was used for the analysis. The band intensities were analyzed using ImageJ software (National Institute of Health, Bethesda, MD, USA), as previously described [35 (link),46 (link),64 (link),82 (link),83 (link)].