Fig. 5.

STK25 regulates YAP phosphorylation in response to physiologic stimuli. a Immunoblot and quantification of YAP phosphorylation in HEK293A cells grown to low confluence or high confluence after transfection with the indicated siRNA (n = 3 biological replicates; *p < 0.05, unpaired t-test). b Cellular proliferation curves of control HEK293A clones and STK25 KO clones over the indicated time periods (n = 3 replicates per cell line; ***p < 0.001, ****p < 0.0001, two-way ANOVA with Tukey’s post-hoc test). c Representative immunoblot and quantification of YAP phosphorylation in IMR90 fibroblasts transfected with the indicated siRNA and held in suspension for the indicated time periods. d Quantification of the percentage of cells remaining in S/G₂ phase following prolonged contact inhibition in hTERT-RPE-1-FUCCI cells transfected with the indicated siRNA (n = 4 biological replicates; **p < 0.01, ****p < 0.0001, one-way ANOVA with Dunnett’s post-hoc test). e Cytokinesis failure was pharmacologically induced in hTERT-RPE-1 cells to generate binucleated tetraploid cells, and the percentage of EdU-positive tetraploid cells following siRNA transfection was quantified. Cells were stained for DNA (white) and EdU incorporation (green). Scale bar, 20 µm. f Quantification of the percentage of EdU-positive binucleated tetraploid cells following transfection with the indicated siRNA. TP53 siRNA served as positive control (n = 4 biological replicates; *p < 0.05, one-way ANOVA with Dunnett’s post-hoc test). g Immunoblot and quantification of LATS1 hydrophobic motif (P-LATS-HM) and activation loop (P-LATS-AL) phosphorylation in either control HEK293A stably expressing Cas9 and a non-targeting sgRNA or STK25 KO 293A stably expressing Cas9 together with sgRNA 1 grown to confluence (n = 3 biological replicates; *p < 0.05, **p < 0.01, paired t-test). All data are presented as mean ± SEM