Extended Data Fig. 9: Targeting ATR rescues the YAP/TAZ activity.
a Quantifications of immunostaining for YAP/TAZ in WT and MLL4P4093X MSCs. Box plots indicate the median (middle line), the first and third quartiles (box), and the 10th and 90th percentile (error bars). The number of analyzed nuclei is reported in figure as n; unpaired two-tailed Student’s t-test was applied for statistical analysis (****P<0.0001). b Western Blot analysis of YAP, TAZ and phosphorylated YAP (Ser127) in WT and MLL4Q4092X MSCs; GAPDH was used as loading control. c Representative images of the distribution of ATR in WT and MLL4Q4092X MSCs. Scale bar, 20 µm. d Representative images and quantifications of immunostaining for YAP/TAZ in WT and MLL4Q4092X MSCs untreated or treated with different concentrations of the ATR inhibitor VE-822. Scale bar, 20 µm. Data are represented as box plots, indicating the median (middle line), the first and third quartiles (box), and the 10th and 90th percentile (error bars). The number of analyzed nuclei is reported in figure as n; unpaired two-tailed Student’s t-test was applied for statistical analysis (****P<0.0001). e Representative images and quantification of phH2A.X in WT and MLL4Q4092X MSCs. Scale bar, 20 µm. Box plots indicate the median (middle line), the first and third quartiles (box), and the 10th and 90th percentile (error bars) of the phH2A.X distribution per cell. The number of analyzed nuclei is reported in figure as n; unpaired two-tailed Student’s t-test was applied for statistical analysis. f Volcano plot of differentially expressed genes between WT and MLL4Q4092X MSCs. Vertical blue lines indicate the chosen cutoff (−2>fold change>2), horizontal red line indicates Wald test-derived p-value cutoff of 0.05. g Heat map of k-means clustering analysis of expressed genes in WT MSCs untreated or treated with ATR inhibitor at the indicated time points. h Venn diagram showing the overlap between the ATR responsive genes identified in WT and MLL4Q4092X MSCs. i qRT-PCR of the indicated genes in WT, MLL4Q4092X (i) and MLL4P4093X MSCs (k) untreated or treated with ATR inhibitor at the different time points. The expression level was normalized on GAPDH level. Data are means + SEM (n=3 independent experiments); unpaired two-tailed Student’s t-test was applied for statistical analysis (***P<0.001).
