Fig. 5.
p53 favors the functional recovery of CPCs from oxidative stress in vitro. (a) Western blotting of p16INK4a at baseline, after Doxo-pulse and following recovery of WT-CPCs (WT) and p53-tg-CPCs (p53-tg); n = 3 in all cases. Optical density data are mean ± SD. *p < 0.05 vs. WT-Control. **p < 0.05 vs. WT-Doxo-pulse. ***p < 0.05 vs. WT-recovery. (b) p16INK4a labeling (upper left panel, yellow) of WT-CPCs exposed to Doxo. Nuclei are stained by DAPI (upper right panel, blue). Phalloidin (lower left panel, white). Merge of p16INK4a, DAPI and phalloidin (lower right panel). Scale bar, 50 μm. Fraction of p16INK4a-positive WT-CPCs and p53-tg-CPCs at baseline, following Doxo-pulse and after recovery; n = 3 in all cases. Data are mean ± SD. *p < 0.05 vs. WT-Control. **p < 0.05 vs. WT-Doxo-pulse. ***p < 0.05 vs. WT recovery. †p < 0.05 vs. p53-tg control. ‡p < 0.05 vs. p53-tg Doxo-pulse. (c) Number of DDR foci in WT-CPCs and p53-tg-CPCs at baseline, after Doxo-pulse and following recovery; n = 3 in all cases. For statistics see panel B. (d) Nucleoids in WT-CPCs and p53-tg-CPCs at baseline, following Doxo-pulse and after recovery are stained with Vista green dye (green). Comets are apparent in Doxo-pulse and after recovery of WT-CPCs, while intact DNA is noted in p53-tg-CPCs after recovery. (e) Damaged DNA in nuclei of WT-CPCs and p53-tg-CPCs at baseline, after Doxo-pulse and following recovery; n = 3 in all cases. For statistics see panel B. (f) Fraction of Ki67-positive WT-CPCs and p53-tg-CPCs following 24, 48 and 72 h recovery period; n = 3 in all cases.*p < 0.05 vs. 24 h. **p < 0.05 vs. 48 h.