Rabbit anti mlc2v

Cardiac tissues were fixed with 4% paraformaldehyde (Nacalai Tesque). Experiments were performed on paraffin sections after antigen retrieval with citrate buffer (pH 6.0) at 121°C for 20 min. The tissues were incubated with primary antibodies including mouse or rabbit anti-cTnT (1:200 dilution; Abcam, Cambridge, UK), rabbit anti-vimentin (1:200, Abcam), mouse anti-sarcomeric α-actinin (1:400; Sigma-Aldrich), rabbit anti-connexin43 (1:100; Abcam), mouse anti-MLC2a (1:100; Synaptic Systems, Goettingen, Germany), rabbit anti-MLC2v (1:200; Proteintech, Rosemont, IL, USA), mouse anti-FN (1:200; Abcam), rabbit anti-laminin (1:30; Sigma-Aldrich), and rabbit anti-collagen type I (1:500; Abcam). The endothelial cells were immunostained with mouse anti-CD31 antibody (1:200; Dako, Glostrup, Denmark). The tissues were permeabilized with 0.2% Triton X (Sigma-Aldrich), and then non-specific reactivity was blocked with 1% BSA. The tissues were labeled by primary antibodies. Secondary antibodies (1:200) such as Alexa Fluor 488- or Alexa Fluor 546-conjugated goat anti-mouse or anti-rabbit IgG (H+L) (Thermo Fisher Scientific) were added to the tissues. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) (Life Technologies) and observed by confocal laser scanning microscopy (FLUOVIEW FV10i; Olympus, Tokyo, Japan).