Mouse monoclonal anti tgf β1 antibody

Sample collection and western blotting were performed as described previously [20 (link)]. Rabbit polyclonal anti-Sirt1 antibody (Sigma-Aldrich), rabbit monoclonal anti-p16^INK4a antibody (Abcam, Cambridge, UK), mouse monoclonal anti-GAPDH antibody (Sigma-Aldrich), mouse monoclonal anti-α-SMA antibody (Sigma-Aldrich), mouse monoclonal anti-TGF-β1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit monoclonal anti-phosphorylated Smad2 (p-Smad2) antibody (Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal anti-Smad2 antibody (Cell Signaling Technology), mouse monoclonal anti-α-tubulin antibody (Sigma-Aldrich), rabbit monoclonal anti-CD163 antibody (Abcam), and rabbit polyclonal anti-CD68 antibody (Abcam) were used as primary antibodies. Horseradish peroxidase-conjugated goat anti-rabbit immunoglobulin G (Dako, Glostrup, Denmark) or goat anti-mouse immunoglobulin G (Dako) were used as secondary antibodies. SuperSignal West Dura or Pico system (Thermo Fisher Scientific, Rockford, IL, USA) was used to detect signals. The intensity of each band was analyzed by ImageJ software (version 1.47v; National Institutes of Health) and standardized by the level of either GAPDH or α-tubulin.

As primary antibodies, we used a rabbit monoclonal anti-HMGB1 antibody (ab79823; Abcam, Cambridge, UK), rabbit monoclonal anti-IL-18 antibody (ab223293; Abcam), rabbit monoclonal anti-IFN-γ antibody (ab133566; Abcam), mouse monoclonal anti-α-SMA antibody (A-2547; Sigma-Aldrich; St. Louis, MO), mouse monoclonal anti- TGF-β1 antibody (sc-130348; Santa Cruz Biotechnology, Santa Cruz, CA), mouse monoclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody (G8795; Sigma-Aldrich), rabbit monoclonal anti-p-Smad2 antibody (#3108; Cell Signaling Technology, Danvers, MA), mouse monoclonal anti-Smad2 antibody (#3103; Cell Signaling Technology), rabbit polyclonal anti-Col-I antibody (ab6308; Abcam), rabbit polyclonal anti-Col-III antibody (ab7778; Abcam), rabbit polyclonal anti-CD3 antibody (IR503; Dako, Santa Clara, CA), rabbit polyclonal anti-CD68 antibody (ab125212; Abcam), rabbit polyclonal anti-CD206 antibody (ab64693; Abcam) and rabbit monoclonal anti CD163 antibody (ab182422; Abcam).

Sample collection and western blotting were performed according to previously described methods [26 (link)]. Mouse monoclonal anti-α-SMA antibody (Sigma-Aldrich), mouse monoclonal anti-TGF-β1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse monoclonal anti-GAPDH antibody (Sigma-Aldrich), rabbit monoclonal anti-p-Smad2 antibody (Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal anti-Smad2 antibody (Cell Signaling Technology), and mouse monoclonal anti-α-Tubulin antibody (Sigma-Aldrich) were used as primary antibodies. Horseradish peroxidase-conjugated goat anti-rabbit immunoglobulin G (Dako, Glostrup, Denmark) or goat anti-mouse immunoglobulin G (Dako) was used as secondary antibodies. SuperSignal West Dura or the Pico system (Thermo Fisher Scientific, Rockford, IL, USA) were used to detect signals. The intensity of each band was measured by ImageJ software (version 1.47v; National Institutes of Health) and normalized to the level of either GAPDH or α-tubulin.