Figure 5. NDR1 impairs TGFβ-phosphorylation of Smad2.

A. Lysates of untreated or 48h-TGFβ-incubated NMuMG cells transfected with an expression plasmid encoding wild type or kinase-inactive NDR1, or transfected with the vector control were subjected to immunoblotting using an antibody that recognizes Smad2 when phosphorylated specifically at the TGFβ-induced sites (pSmad2) or an antibody that recognizes Smad2 regardless of its phosphorylation status (Smad2) or with an actin antibody, the latter serving as a loading control. B. Actin-normalized TGFβ-phosphorylated Smad2 was expressed relative to the actin-normalized total Smad2. Data are presented as the mean+SEM (n = 3) ratio of TGFβ-phosphorylated Smad2 to total Smad2. Statistical significance between TGFβ-induced phosphorylation of Smad2 in the vector control cells and each of the wild type and kinase-inactive NDR1-expressing cells is indicated (ANOVA).