Figure 9

TGFβ-induced changes in p65/RelA phosphorylation are dependent on Par6 and TβRI activation. Subconfluent monolayers of the indicated NMuMG-derived cell lines were treated for 48 (A, B) or 144 (C, D) hours with control media (DMSO alone), TGFβ1 (TGFβ, 5 ng/ml), the TβRI inhibitor SB-431542 (SB, 10 μM) or TGFβ + SB. The level of phosphorylated p65/RelA (pRelA, S536), native p65/RelA and α-tubulin (loading control) was examined by immunoblotting. Graphs in B and D represent the average relative p65/RelA phosphorylation at S536 as determined by band densitometry for 3 independent experiments. Relative p65/RelA phosphorylation was calculated as the ratio of phosphorylated p65/RelA to native p65/RelA. The value for DMSO control was set to 1 for each cell line. Two-way ANOVA for all cell lines and treatments was significant (p < 0.01) only for the 144-hour treatment. Bonferroni post test compared differences between control and treatment for each cell line, *p < 0.05, **p < 0.001.