Catalog no ab42476

Tissues and cells were lysed in RIPA lysis buffer (Catalog No. P0013B, Beyotime). The lysis was centrifuged, and the supernatant was collected for Western blot analysis. Samples of protein were resolved on SDS‐PAGE and transferred onto PVDF membranes (Catalog No. ISEQ00010, Millipore). The membranes were incubated with the following primary antibodies overnight at 4°C: anti‐Hic‐5 (Catalog No. ab42476, 1:1000, abcam), anti‐α‐SMA (Catalog No. ab5694, 1:500, abcam), anti‐Col1a1 (Catalog No. GB11022, 1:1000, Servicebio), anti‐IL‐6 (Catalog No. GB11117, 1:1000, Servicebio), anti‐NF‐κB/p65 (Catalog No. 10745‐1‐AP, 1:1000, Proteintech), anti‐Vimentin (Catalog No. GB11192, 1:1000, Servicebio) and anti‐GAPDH (Catalog No. 60004‐1‐Ig, 1:5000, Proteintech). Then, the membranes were incubated with horseradish peroxidase‐conjugated secondary antibodies for 1 hour at room temperature. The following secondary antibodies conjugated to horseradish peroxidase: Affinipure goat antimouse IgG (Catalog No. SA00001‐1, 1:5000, Proteintech) and Affinipure goat anti‐rabbit IgG (Catalog No. SA00001‐2, 1:5000, Proteintech).

Antigen retrieval was performed by heating slides in the microwave for 20 minutes in 0.01 mol/L citrate buffer (pH 6.0), and 3% hydrogen peroxide was added for 10 minutes to quench peroxidase activity. Sections were treated with normal goat serum, followed by incubation overnight with anti‐Hic5 antibody (Catalog No. ab42476, 1:100, abcam) or anti‐α‐SMA antibody (Catalog No. ab5694, 1:100, abcam) at 4°C. Sections were then rinsed with phosphate‐buffered saline (PBS), incubated with secondary antibody for 1 hours, stained with diaminobenzidine and counterstained with haematoxylin. Sections were dehydrated and sealed, then examined using a microscope (Olympus).