Fig. 6.

ES2 disrupts FOXO4 foci and localizes to FOXO4 foci. a, Representative images of FOXO4 and TP53BP1 (53BP1) in A375 cells treated as described in Fig. 3a. Scale bar is 10 µm. b, Scatterplot of the FOXO4 nuclear intensity vs. the DNA content (DAPI intensity) of individual cells either mock-treated (dividing) or doxorubicin-treated (senescent) A375 cells. Red dots indicate cells treated with doxorubicin. Units of fluorescence intensity are arbitrary. c, Box and whisker plots of the FOXO4 foci/nucleus in A375 cells treated as described in a. **** p-value < 0.0001. d, Representative images of FOXO4 and active caspase-3/7 in senescent A375 cells treated with ES2 8 µM for 4 h. e, Bar graphs of the median FOXO4 foci/nucleus and median FOXO4 nuclear intensity in A375 cells treated as described in e. unt (untreated). Error bars denote the 95% confidence interval. **** p-value < 0.0001. f, Plots of median fluorescence intensity of ES2-BODIPY at 2 h after addition of labeled ES2 to dividing or senescent A375 cells. g, Representative images of A375 senescent cells 2 h after addition of ES2-BODIPY showing the overlay of FOXO4 (purple) and ES2 (green) outside of the nucleus in dying cells. The nucleus is stained with DAPI (blue) and outlined. Scale bar = 10 µm.