Immunohistochemical Staining of Lung Sections

Immunohistochemical staining was performed on paraffin embedded lung sections. The sections were deparaffinized with xylene and prepared for staining as described previously. Sections were blocked with 10% goat serum in PBS for 1 h, followed by incubation with the following primary antibodies at 4 °C for 12 h: rabbit anti-mouse α-SMA (Abcam, ab124964, dilution 1:500) and rabbit anti-mouse COL1A (Abcam, ab260043, dilution 1:500). After washing, sections were incubated with biotin-conjugated goat anti-rabbit IgG and avidin–biotin peroxidase complex (DBA) at room temperature for 2 h, and photographed using a light microscope (Motic, China) at the magnification of × 100. To assess nonspecific staining for α-SMA and COLA1, alternate sections from each experimental condition were also incubated with primary or secondary antibody only [20 (link)].