Anti tgf β1

For indirect immuno-fluorescence (IIF), 1.5 × 10⁴ cells were incubated with anti-collagen type I (COL1A1), anti-cellular fibronectin, and anti-α-SMA followed by goat anti-mouse FITC-conjugated antibody. Nuclei were visualized using Hoechst 33342 (all from Sigma-Aldrich, Milano, Italy). For immuno-cytochemistry (ICC), 1.5 × 10⁴ cells were incubated with anti-TGF-β1 (Sigma-Aldrich) and then immune-stained using the streptavidin–biotin–peroxidase technique (Dako Cytomation, Milano, Italy) and incubated with 3,3-diaminobenzidine. Slides were counterstained with Mayer’s hematoxylin. To quantify the expression of the proteins, the percentage of area occupied by the protein inside the cells was calculated using Fiji-ImageJ software (available online at https://imagej.net/Fiji) [22 (link)].

Sch C, purity: 99.76%, Chengdu Pufei De Biotech Co., Ltd., Chengdu, China; BLM, Nippon Kayaku Co., Ltd. Japan; JNK inhibitor (SP600125), Selleck China Inc. (Shanghai, China); TGF-β₁, Sigma-Aldrich Inc. (Shanghai, China); The primary anti bodies are both from Abclonal: anti-α-SMA, anti-FN, anti-Collagen type I (Col 1A1), anti-Collagen type III (Col 3A1), anti-LOX, anti-TGF-β₁, anti-Smad2, anti-Smad3, anti-phosphorylation-Smad2 (p-Smad2), anti-phosphorylation-Smad3 (p-Smad3), anti-JNK, anti-phosphorylation-JNK (p-JNK), anti-MMP2, anti-MMP9, anti-TIMP1, and anti-β-actin. The dilution ratio of all antibodies was 1:1000.