Smad2 3 sirna

mCherry-EGFP-LC3 lentivirus vectors, ATG5-siRNA lentivirus vectors and BECN1-siRNA lentivirus vectors were purchased from GeneChem Technology (Shanghai, China). The control siRNA, Smad2/3 siRNA and c-Jun siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). 3-methyladenine (3-MA, M9281) was purchased from Sigma-Aldrich. Bafilomycin A1 (Baf A1; S1413) was purchased from Selleck. TGF-β1 ELISA reagents (MB100B) and TGF-β2 ELISA reagents (MB200) were purchased from R&D systems. TGF-β3 ELISA reagents (E-EL-M1192) was purchased from Elabscience Biotechnology. Recombinant human TGF-β1 (100-21) and recombinant human TGF-β3 (100-36E) were purchased from PeproTech.

TGF-β was purchased from PeproTech, Inc. FXR1 (cat. no. 67813-1-Ig, 1:10,00 or 1:100), Ki67 (27309-1-AP; 1:100) and β-actin (cat. no. 66009-1-Ig, 1:10,00) antibodies were purchased from ProteinTech Group, Inc. The FXR1 antibody was diluted 1:1,000 in the western blotting assay and 1:100 in immunohistochemistry assays. SMAD2/3 (cat. no. # 8685S, 1:1,000), N-Cadherin (cat. no. # 4061, 1:1,000), and slug (cat. no. 9585S, 1:1,000) antibodies were purchased from Cell Signaling Technology, Inc. The secondary antibodies used were HRP-conjugated Affinipure Goat Anti-Rabbit (1:2,000 or 1:50; cat. no. SA00001-2) and HRP-conjugated Affinipure Goat Anti-Mouse (1:2,000; cat. no. SA00001-1) (both from ProteinTech Group, Inc.). HRP-conjugated Affinipure Goat Anti-Rabbit was diluted 1:2,000 in the western blotting assay and 1:50 in immunohistochemistry assays. SMAD2/3 siRNA was purchased from Santa Cruz Biotechnology, Inc. (cat. no. sc-37238). shRNA was purchased from Public Protein/Plasmid Library. The sequences of the FXR1 knockdown shRNAs were: shFXR1#1, 5′-GCTAGAGGTTTCTTGGAATTT-3′; shFXR1#2, 5′-CGCCAGGTTCCATTTAATGAA-3′; and negative control (shNC), 5′-GTTCTCCGAACGTGTCACGTT-3′. The FXR1 expression plasmid with a 3′FLAG tag was purchased from Public Protein/Plasmid Library.