Anti αsma antibody

Specimens were stained with hematoxylin and eosin (H&E) and Masson’s trichrome, and immunostained for F4/80 or alpha smooth muscle actin (αSMA), as previously reported [15 (link), 16 ]. In brief, heart samples were harvested from PBS- or talc-injected mice and fixed with 4% paraformaldehyde overnight. Tissues were then embedded in paraffin and sectioned into 10 μm slices along the cephalocaudal axis. For avidin-biotin complex (ABC) immunostaining, sections were incubated with anti-F4/80 antibody (1/100 dilution; Clone# A3–1, Abcam, Cambridge, MA) or anti-αSMA antibody (1/100 dilution; Clone# SPM332, Novus Biologicals, Littleton, CO, USA) overnight, followed by horseradish peroxidase-labelled secondary antibody (Nichirei Bioscience, Tokyo, Japan) for 1 h. Visualization of F4/80 and αSMA was achieved via 3,3′-diaminobenzidine tetrahydrochloride (DAB) reaction. Images were taken using a microscope (Olympus, Tokyo, Japan).

Anti-CD31 rabbit polyclonal antibody was obtained from Spring Biosciences (Catalogue number E11114; Pleasanton, CA, USA). Anti-αSMA antibody was purchased from Novus Biologicals (Clone SPM332; Littleton, CO, USA). Anti-CD68 (Clone PG-M1) and anti-CD34 (Clone QBEnd10) antibodies were obtained from DAKO (Carpinteria, CA, USA). Anti-fibrinogen β-chain and anti-plasminogen antibodies were from Sigma-Aldrich. Alexa Fluor 488-conjugated goat anti-mouse IgG antibody and Alexa Fluor 567-conjugated goat anti-rabbit IgG antibody were purchased from Life Technologies Japan. Bio-Safe Coomassie Blue was purchased from Bio-Rad Laboratories.