Smad2 3 antibody sampler kit

HEK293 cells and mouse fibroblast NIH3T3 cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). Human primary lung fibroblast cells (HFL) were established by Dr. Reynold Panettieri’s laboratory from patients with brain-related disease but no history of pulmonary fibrosis^{40 (link)}. The cells were frozen at an early passage and cultured for a maximum of fifteen passages. The cells were maintained at 37 °C with 5% CO₂ in Dulbecco’s Modified Eagle’s Medium/Ham’s Nutrient Mixture F-12 (DME/F12, Life Technologies/Gibco, Grand Island, NY) supplemented with 10% fetal bovine serum (GE Healthcare Bio-Sciences, Pittsburgh, PA, USA), 100 U/mL penicillin G and 100 μg/mL streptomycin (Life Technologies/Gibco). Smad3 inhibitor SIS3 and p38MAPK inhibitor SB203580 were purchased from (AdooQ BioScience, Irvine, CA, USA). Recombinant human TGF-β1 and the Smad2/3 Antibody Sampler Kit were obtained from Cell Signaling Technology (Danvers, MA, USA). The mouse anti-α-SMA monoclonal antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA). The rabbit anti-TGFBR1 antibody was obtained from Merck Millipore (Billerica, MA, USA). The mouse anti-CTGF monoclonal antibody, mouse anti-GAPDH monoclonal antibody were obtained from Proteintech (Chicago, IL, USA). The GAPDH levels served as internal controls.