Figure 2

TGFB1 stimulates 3TP-luc activity and Smad2/3 phosphorylation in NIH3T3 cells. A) Murine fibroblast NIH3T3 cells were seeded in 24-well plates and transfected with the TGFB responsive promoter-reporter 3TP-luc. Following transfection, cells were treated with 4–400 pM TGFB1 for approximately 24 hours. Luciferase assays were performed as described. Data points reflect mean (+/- SEM) fold-change in luciferase activity from the control condition (0 pM, not pictured) in two experiments performed in triplicate (n = 6). The data are presented on a log-linear plot. Points with letters differed from control and points with different letters differed from one another. B) NIH3T3 cells seeded in 6-well plates were treated with vehicle (control) or 400 pM TGFB1 for 1 hour. Immunoblots on whole cell lysates were performed as described in the legend to Figure 1, except in the bottom blot an anti-Smad3 antibody was used in place of anti-Smad2/3. Treatments were performed in duplicate. Numbers at the left are molecular weight standards in kDa.