Anti mef2d antibody

ASM cells were harvested, and then, nuclear/cytosol fractionation was performed with a commercially available kit (BioVision), according to the manufacturer’s instructions. The nuclear/cytosol fraction was then sampled, and proteins were separated by SDS-polyacrylamide gel electrophoresis, transferred to nitrocellulose membranes, and immunoblotted with anti-Mef2D antibody (BD Biosciences) and anti-cleaved Caspase-7 antibody (Cell Signaling Technology). Anti-H3 histone antibody (Cell Signaling Technology) was used as loading controls for nuclear fractions.

All cell culture reagents were purchased from Corning Life Sciences. Antibodies against β-tubulin, Yy1, and RhoA were purchased from Santa Cruz Biotechnology. Antibodies against Mlc; phospho-Mlc (Ser19); total Caspase-7 and Caspase-9; cleaved Caspase-3, Caspase-7, Caspase-8, and Caspase-9; cleaved PARP; Calpain-1; Calpain-2; and H3 histone were purchased from Cell Signaling Technology. Anti-Mef2D antibody was purchased from BD Biosciences. Horseradish peroxidase (HRP)-conjugated goat anti-mouse secondary antibody was purchased from Bio-Rad, and HRP-conjugated goat anti-rabbit antibody was purchased from Cell Signaling Technology. Anti-α smooth muscle actin (α-SMA) antibody was purchased from R&D Systems. All chemicals were purchased from Sigma-Aldrich. Reagents for quantitative polymerase chain reaction (qPCR) were purchased from Life Technologies. The mRNA Isolation Kit was purchased from QIAGEN.