Anti col1a1 72026

The antibodies used in the present study were as follows: anti-COL1A1 (72026) and anti-PPARγ (C26H12) from Cell Signaling Technologies (Boston, USA); and anti-ITIH4 (ab180139) from Abcam (Cambridge, UK). Secondary antibodies were either goat Alexa 488 (A-11008) or 594 (A-11012) obtained from ThermoFisher (Oxford, UK). Lipopolysaccharide (LPS, L2630) was purchased from Sigma (Steinheim, Germany).

HSCs cocultured with ADSC were fixed with 4% paraformaldehyde solution for 1 h, and permeabilization was performed with 2% bovine serum albumin (BSA, Sigma-Aldrich) solution with 0.1% Triton X-100 for 20 min. This was followed by overnight incubation at 4 °C with specific antibodies: anti-α-SMA (sc-53015, Santa Cruz Biotechnology, Dallas, TX, USA), anti-SMAD2/3 (5678S, Cell Signaling), anti-COL1A1 (72026, Cell Signaling), and antifibronectin (sc-69681, Santa Cruz Biotechnology). Samples were incubated with specific anti-rabbit/anti-mouse secondary antibodies (Thermo Fischer Scientific) conjugated with AF 546 and AF 488, respectively, and further were stained with Hoechst 33342 (Sigma-Aldrich) for 5 min to visualize the nuclei in blue. Samples were visualized by fluorescence microscopy (IX-73 Olympus).