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. 2008 Dec 18;28(2):88–98. doi: 10.1038/emboj.2008.266

Figure 3.

Figure 3

Depletion of Smad1/5 but not Smad2/3 inhibits TGFβ-stimulated migration in 4T1 cells. Puromycin-selected cells expressing empty vector (pSicoR-puro), Smad1 targeting shRNA (1A or 1B), or Smad5 targeting shRNA (5B or 5C) were subjected to (A) western blot analysis after 45 min. TGFβ treatment or (B) transwell migration for 20 h in the presence or absence of TGFβ. Asterisk indicates detection of Smad1 by Smad5 antibody. Puromycin-selected, GFP+ cells expressing empty vector (pSicoR-puro+GFP), 1A+5C shRNA, 5B+1B shRNA, and 1B+5C shRNA were subjected to (C) western blot analysis after 45 min. TGFβ treatment or (D) transwell migration for 20 h in the presence or absence of TGFβ. Two-tailed Student's t-test was performed (**P=0.05, ***P=0.01, ****P=0.11). Cells expressing control or S3+S2 shRNA were subjected to (E) western blot analysis after 45 min. TGFβ treatment or (F) transwell migration for 20 h in the presence or absence of TGFβ. Migration assays represent the average of at least three independent experiments.