Renca

The human renal cell carcinoma cell lines, 786-O and ACHN, were purchased from the Shanghai Institute of Biochemistry and Cell Biology, China Academy of Sciences (Shanghai, China) and cultured in DMEM and RPMI-1640 Medium, containing 10% fetal bovine serum (FBS), 1% penicillin–streptomycin, at a 37°C incubator with 5% CO₂. The murine renal adenocarcinoma cell line Renca was purchased from Cobioer Biosciences and cultured in RPMI-1640 supplemented with 10% FBS, 100 units/ml penicillin, 100 μg/ml streptomycin, 2 mM L-glutamine, 1 mM NEAA, and 1 mM sodium pyruvate solution. Most of the experiments were carried out when cells were grown to 70% confluence. Dulbecco’s Modified Eagle Medium and RPMI-1640 Medium were purchased from Gibco. DMSO was purchased from SIGMA. PFD was from Selleckchem (#S2907). Recombinant human TGF-β1 (#100-21, Peprotech) was reconstituted in 4 mM HCl containing 1 mg/ml bovine serum albumin (BSA). Anti-E-cadherin mAb (#AB40860, Absci), anti-N-cadherin mAb (#AB21474, Absci, China), anti-c-Ski mAb (ab131134, Abcam), anti-Smad2/3 mAb (#5678, CST), anti-phosphorylated Smad2 mAb (#8828, CST) and anti-GAPDH mAb (#sc-32233, Santa Cruz) were used for Western blot. PE/Cy7 anti-mouse CD45 Antibody (#147704, Biolegend), APC anti-mouse/human CD11b Antibody (#101212, Biolegend) and FITC anti-mouse Gr1 Antibody (#108405, Biolegend) were used for flow cytometry.