Figure 2.

Verteporfin treatment results in dramatically reduced levels of YAP and TAZ. NRK49F fibroblasts were treated with or without 250 nmol/L verteporfin for a total of 12 hours. TGF-β (10 ng/ml) was added for the indicated times. In A, after treatment with verteporfin and TGF-β as described above, NRK49F fibroblasts were stained with an antibody directed against YAP/TAZ (green) and counterstained with DAPI (blue) to identify nuclei. Representative images are shown showing loss of whole–cell YAP/TAZ staining even before TGF-β stimulation but no change in the predominantly nuclear localization of YAP/TAZ. Scale bar, 15 μm. (B) The percentage of cells with predominantly nuclear YAP/TAZ staining was quantified at each time point. A minimum of 50 cells was counted per replicate (four replicates per condition). In C, after treatment with verteporfin and TGF-β, lysates were immunoblotted for YAP, TAZ, and GAPDH. Representative blots are shown in C, and quantification of relative levels of (D) YAP (n=3 per condition) and (E) TAZ (n=5 per condition) are also shown. One-way ANOVA with post hoc Fisher least significant difference was performed. AU, arbitrary unit; VP, verteporfin. *P<0.05.