Monoclonal anti α tubulin sc 23948

The polyclonal anti-SMAD4 (9515) antibody was purchased from Cell Signaling Technology (Danvers, MA). Monoclonal anti-α-tubulin (sc-23,948) and polyclonal anti-CTGF (sc-14,939) antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Horseradish peroxidase (HRP)-conjugated goat anti-mouse and goat anti-rabbit immunoglobulin G were obtained from Bio-Rad Laboratories (Hercules, CA). HRP-conjugated donkey anti-goat immunoglobulin G was obtained from Santa Cruz Biotechnology. Recombinant human BMP2, dorsomorphin dihydrochloride (dorsomorphin), and dorsomorphin homolog 1 (DMH-1; 4-[6-[4-(1-methylethoxy) phenyl] pyrazole [1, 5-a] pyrimidin-3-yl]-quinoline) were obtained from R&D Systems (Minneapolis, MN). CTGF overexpression and CONTROL vectors were obtained from Gene Copoeia (Rockville, MD). In this study, two specific protein kinase inhibitors DMH-1 (an inhibitor of ALK2/3) and dorsomorphin (an inhibitor of ALK2/3/6) were used to conduct the loss of function experiment by blocking the downstream signaling pathways of ALK2/36. Specifically, the cells were pretreated with DMH-1 (0.25 μM) or dorsomorphin (10 μM) for 1 h, and cells were then treated with a vehicle control or 50 ng/ml BMP2 for 6 h (for examining mRNA levels) or 12 h (for examining protein levels).

The polyclonal anti-Sprouty2 antibody (#S1444) was obtained from Sigma. The monoclonal anti-E-cadherin antibody (#610181) was obtained from BD Biosciences (Mississauga, ON). The polyclonal anti-EGFR (#sc-03) and monoclonal anti-α-tubulin (#sc-23948) antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). The polyclonal anti-ERK1/2 (#9102), anti-Slug (#9585) and monoclonal anti-Snail (#3895) antibodies were obtained from Cell Signaling Technology (Danvers, MA). Horseradish peroxidase-conjugated goat anti-mouse IgG and goat anti-rabbit IgG were obtained from Bio-Rad Laboratories (Hercules, CA). AG1478 and LY294002 were obtained from Sigma. U0126 was obtained from Calbiochem (San Diego, CA). Recombinant human AREG was purchased from R&D Systems (Minneapolis, MN).

Polyclonal anti-CTGF (#sc-14939) and monoclonal anti-α-tubulin (#sc-23948) antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Polyclonal anti-SMAD4 (#9515), anti-TGF-β receptor type I (#3712), anti-TGF-β receptor type II (#3713), anti-caspase-3 (#9662), monoclonal anti-SMAD2 (#3103) and anti-SMAD3 (#9523) antibodies were obtained from Cell Signaling Technology (Danvers, MA). Horseradish peroxidase-conjugated goat anti-mouse IgG and goat anti-rabbit IgG were obtained from Bio-Rad Laboratories (Hercules, CA). Horseradish peroxidase-conjugated donkey anti-goat IgG was obtained from Santa Cruz Biotechnology. Recombinant human TGF-β1 was obtained from R&D Systems (Minneapolis, MN). SB431542 was obtained from Sigma.