Ab15007

HSCs were co-cultured with either SFM, hBM-MSCs, or hBM-MSCs-Ex (5 ng/ml) for 48 h before samples were collected for protein extraction. Liver tissue was collected from each treatment group (liver fibrosis, hBM-MSCs, and hBM-MSCs-Ex group) for protein extraction. Protein samples were mixed with SDS sample buffer and heated to 95 °C for 10 min, followed by separation on SDS-polyacrylamide gels. Resolved proteins were electro-blotted onto nitrocellulose membrane and probed with antibodies against PPARγ (ab 23673), Wnt3a (ab 248472), Wnt10b (ab70816), β-catenin (ab32572), WISP1 (ab50041), Cyclin D1 (ab16663), α-SMA (ab5694), Collagen I (ab138492), and GAPDH (ab 8245) overnight at 4 °C (1:1000 dilution, Abcam, Cambridge, UK). Nitrocellulose membranes were then incubated with a secondary antibody, HRP-conjugated goat anti-rabbit IgG (ab15007), at room temperature for 2 h, and visualized by chemiluminescent detection according to the manufacturer’s instructions (Immobilon western chemiluminescent HRP substrate, Millipore).

HSCs were co-cultured with either PBS, hUCMSC secretome, or hFSSC secretome (5 ng/ml) for 48 h before samples were collected for protein extraction. Protein samples were mixed with SDS sample buffer and heated to 95 °C for 10 min, followed by separation on SDS-polyacrylamide gels. Resolved proteins were electro-blotted onto nitrocellulose membrane and probed with antibodies against TGF-β1 (ab92486), Smad2 (ab40855), Smad3 (ab40854), Smad7 (ab216428), Collagen I (ab90395), and β-actin (ab5694), (1:1000 dilution, Abcam, Cambridge, UK) overnight at 4 °C (1:1000 dilution, Abcam, Cambridge, UK). Nitrocellulose membranes were then incubated with a secondary antibody, HRP-conjugated goat anti-rabbit IgG (ab15007), at room temperature for 2 h, and visualized by chemiluminescent detection according to the manufacturer’s instructions (Immobilon western chemiluminescent HRP substrate, Millipore, Massachusetts, USA).