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. 2022 Jun 29;13:3732. doi: 10.1038/s41467-022-31399-w

Fig. 4. Prolonged MAPK activation leads to cytoskeletal defects and Hippo activation.

Fig. 4

a Representative fluorescence and phase-contrast images from a live-cell video of dox-inducible BRAFV600E Mel-ST cells expressing the chromosome marker H2B-GFP (green) cultured ± dox (scale bar = 25 µm, hh:mm). b Plot of mitotic duration and fate of individually tracked mitoses from (a) (n > 80 mitoses per condition from two independent experiments, graph shows mean ± SEM, dots represent individually tracked mitoses, black P value represent mitotic duration significance, two-tailed unpaired t test, blue P value represent significance for difference in frequency of whole-genome doubling events, two-sided Fisher’s exact test). c Left, IB of indicated dox-inducible BRAFV600E Mel-ST cell lines cultured ± dox for 24 h along with indicated drugs at the following doses: ERKi (20 nM), MEKi-1 (10 µM), MEKi-2 (20 nM); right, intensity quantification of YAP phos-tag (n = 3 independent experiments, graph shows mean ± SEM, one-way ANOVA with multiple comparisons). d Left, IB of dox-inducible BRAFV600E Mel-ST cell lines cultured ± dox for indicated time; right, intensity quantification of YAP phos-tag (n = 3 independent experiments, graph shows mean ± SEM, two-tailed unpaired t test). e Left, representative IB of dox-inducible BRAFV600E Mel-ST cell lines treated ± dox for 24 h or with 1 mM hydroxyurea for 6 h; right, intensity quantification of YAP phos-tag (n = 3 independent experiments, graph shows mean ± SEM, two-tailed unpaired t test). f Representative IB of RhoA-GTP pulldown in indicated dox-inducible BRAFV600E Mel-ST cell line ± dox; right, intensity quantification of RhoA-GTP to total RhoA (n = 3 independent experiments, graph shows mean ± SEM, two-tailed unpaired t test). Source Data are provided as a Source Data file.