a: Partial deletion of LATS1/2 in MC38 diminishes YAP/TAZ phosphorylation in response to actin polymerization inhibitor LatB and energy stress by 2-DG treatment. Wild-type and LATS1/2 partial knockout MC38 clone #5 were seeded on six-well plates and treated with LatB and 2-DG for 1 hour. Cells were collected for determination of YAP/TAZ phosphorylation. l.e., long exposure of films.
b: LATS1/2 deletion abolishes YAP/TAZ phosphorylation in Panc02 cells. Experiments are similar to panel A.
c: LATS1/2 deletion increases the expression of YAP/TAZ target genes Ctgf and Amotl2. mRNA levels of Ctgf and Amotl2 expression were measured by quantitative real-time PCR. Results were from triplicated experiments. The error bars represent s.d. ** p<0.01; *** p<0.001. Student’s t-test was applied.
d-e: LATS1/2 deletion induces constitutive nuclear YAP/TAZ localization. LatB-treated or non-treated MC38 or Panc02 cells were subjected to immunostaining with the YAP/TAZ antibody (green) along with DAPI for DNA (blue). Scale bar, 10 μm.
f: Quantification of data from panels D-E. Ten randomly chosen fields for each treatment were examined. Typically, 100 cells were counted in each field. N and C denote nuclear and cytoplasmic.