Rabbit anti collagen 1

The liver tissues and LX2 cells were washed, homogenized on ice with RIPA buffer (Sigma-Aldrich, Saint Louis, MO, USA), and centrifuged at 8000 rpm for 10 min. The protein concentration in the supernatant was determined using a Bradford assay. Lysates containing equal amounts of protein were separated by SDS-PAGE. The western blot analysis was performed as previously described^{21 (link)}. The following primary antibodies were used in this study: rabbit anti-BCL2L10 (1:500, Origene), rabbit anti-glyceraldehyde phosphate dehydrogenase (GAPDH; 1:8000, Cell Signaling Technology), rabbit anti-cleaved caspase3 (1:400, Affinity), rabbit anti-cleaved caspase9 (1:400, Affinity), rabbit anti-collagen I (1:700, Affinity), rabbit anti-α-smooth muscle actin (α-SMA; 1:1000, Abcam), rabbit anti-TOM20 (1:2000, Affinity), and mouse anti-BCL2L10 (1:400, Abcam). After incubating with fluorescence-conjugated secondary antibodies (1:20000, Rockland Biochemicals), the immunoreactive bands were visualized using an Odyssey Infrared Imaging System (LI-COR Biosciences). For the protein quantification, the bands were scanned and quantified using Image-Pro plus 6.0 software (Datacell, UK), and GAPDH served as an internal control.

Retention of collagen I, collagen IV, fibronectin, and laminin in DTM was studied by immunohistochemistry. Sections were deparaffinized and rehydrated. Endogenous peroxidases were quenched using 0.3% H₂O₂. After blocking with 3% bovine serum albumin (BSA), sections were incubated with primary antibodies at 4°C overnight, followed by incubation using a horseradish peroxidase (HRP)-conjugated secondary antibody at 37°C for 1 h. The peroxidase activity was visualized with 3,3-diaminobenzidine (DAB) for 5 min, and the slides were counterstained with hematoxylin. Normal rabbit immunoglobulin G (IgG) was used for isotype controls. The following antibodies were used: rabbit anticollagen I (Affinity Biosciences, Changzhou, China), rabbit anticollagen IV (Affinity Biosciences, Changzhou, China), rabbit antifibronectin (Affinity Biosciences, Changzhou, China), and rabbit antilaminin (Affinity Biosciences, Changzhou, China).