Immunohistochemical and Fluorescent Staining Procedures

Methods for immunohistochemical staining are described elsewhere 26 (link). Antibodies used for IHC were fibronectin (Abcam), collagen1A1 (Santa Cruz), TGF-β (Abcam), and IL-1β (Santa Cruz). Primary antibody labeling detection was performed after incubation with the appropriate DAB incubated secondary antibody. For fluorescent labeling of Nrf2 nucleus translocation, frozen sections were fixed with 4% PFA for 10 min and permeabilized with 0.1% Triton X-100 for 20 min. After blocking with 5% BSA for 1h, sections were stained with Nrf2 antibody (Abcam), and then stained with secondary antibody, Texas Red (Invitrogen). Nuclei were counterstained with hematoxylin in IHC experiments and with DAPI in SlowFade® Gold Anti-fade Mountant in IF experiments.