Fig. 7.

Tension-dependence of Ajuba and LATS family protein localization correlates with influences of cytoskeletal tension on YAP activity. (A) MCF10A EGFP:LIMD1 cells were plated at low density and grown for 48 h in total. Transgene expression was induced by adding Dox 24 h before treatments. Cells were treated with DMSO (control) or treated with 50 µM blebbistatin for 1 h. After the treatment, cells were stained for YAP (red) and DNA (Hoechst 33342, blue) (B) Western blots of lysates of MCF10A cells treated with DMSO (control) or 50 µM blebbistatin for 1 h, blotted using the indicated antisera. All the blots are from the same experiment and a representative loading control (tubulin) is shown. Histograms show a quantification of the pYAP (S127) to total YAP ratio and pLATS1 (S909 and T1079) to total LATS1 ratio from three biological replicates normalized to the ratio in the control. (C) MDCKIIG EGFP:LIMD1 cells were grown and stained as in A. Images are z-projections and are representative of at least three biological replicates. (D) Western blots of lysates of MDCKIIG cells treated with DMSO (control) or 50 µM blebbistatin for 1 h, blotted using the indicated antisera. Tubulin was used as a loading control. Histograms show a quantification of the pYAP (S127) to total YAP ratio and pLATS1 (S909) to total LATS1 ratio from three biological replicates normalized to the ratio in the control. In all histograms, error bars indicate s.e.m. and P-values less than 0.05 are shown.