Protein Expression Analysis by Western Blotting

The protein expression was assessed by Western blotting. Primary antibodies, i.e., mouse anti-α-SMA antibody (SMA, Sigma-Aldrich, A2547), rabbit anti- MMP-9 antibody (Abcam, Cambridge, UK, ab76003), mouse anti-Toll-like receptor-4 (TLR-4) antibody (Abcam, ab30667), and mouse anti-β-actin antibody for internal control (Sigma-Aldrich, A5411) were reacted with the polyvinylidene fluoride (PVDF) membrane at 4 °C overnight. Subsequently, the corresponding secondary antibodies were immersed at room temperature for 1 h. The protein bands were quantified using Image J software and normalized using individual internal controls for comparison [17 (link),18 (link)].