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. 2017 Jan 31;16:224–237. doi: 10.1016/j.ebiom.2017.01.028

Fig. 3.

Fig. 3

p53 improves the DDR of CPCs. (a) Nuclei from p53-tg-CPCs in the absence (Control) and in the presence of Doxo are stained by DAPI (blue, left panels); immunolabeled γH2A.X is shown in these nuclei (green, right panels). Scale bar: 100 μm. (b) Fraction of γH2A.X-positive CPCs in the absence (control, Ctrl) and following exposure to Doxo (Doxo): Ctrl WT-CPCs (4284 cells from 3 mice); Ctrl p53-tg-CPCs (13,334 cells from 3 mice); Doxo WT-CPCs (3958 cells from 3 mice); and Doxo p53-tg-CPCs (16,496 cells from 3 mice). Data are mean ± SD. (c) γH2A.X (green; left two panels) in nuclei of WT-CPCs and p53-tg-CPCs stained by DAPI (blue). DDR foci are illustrated in the same nuclei following three-dimensional reconstruction by Imaris version 5.5.2 (right two panels). Scale bar: 5 μm. (d) Number of DDR foci counted in nuclei of WT-CPCs and p53-tg-CPCs. In each case, 24–59 γH2A.X positive nuclei from 3 mice were analyzed. (e) Nucleoids of WT-CPCs and p53-tg-CPCs are stained with Vista green dye (green, left panels). Comets are apparent after Doxo (green, right panels). (f) Quantity of damaged DNA in nuclei of WT-CPCs and p53-tg-CPCs at baseline (Control: WT, n = 62 comets from 3 mice; p53-tg, n = 70 comets from 3 mice) and after Doxo (Doxo: WT, n = 76 comets from 3 mice; p53-tg, n = 61 comets from 3 mice). *p < 0.05 vs. WT Ctrl; **p < 0.05 vs. Doxo WT-CPCs; ***p < 0.05 vs. p53-tg Ctrl.