Figure 2

Par6 overactivity sensitizes NMuMG cells to TGFβ-induced apoptosis. Subconfluent monolayers of the indicated NMuMG–derived cell lines were treated with control media (DMSO alone), TGFβ1 (TGFβ; 5 ng/ml), the TβRI/SMAD inhibitor SB-431542 (SB) or TGFβ + SB. The level of cleaved caspase-3 was determined as a read-out of apoptosis after 48 hours (A, B) or 144 hours (C, D) treatment. A and C show levels of cleaved caspase-3 (CC3), uncleaved caspase-3 (C3) and α-tubulin (loading control) as determined by immunoblotting. Cleaved PARP (cPARP) in A served as an additional read-out of apoptosis. Graph in B and D represents the average relative caspase-3 cleavage calculated by band densitometry for 3 independent experiments. Relative C3 cleavage was calculated as the ratio of CC3 to C3. 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 < 0.001). Bonferroni post test compared differences between control and treatment for each cell line, **p < 0.001.