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. 2021 Mar 29;12:649492. doi: 10.3389/fphys.2021.649492

FIGURE 2.

FIGURE 2

DDR is indispensable for regulating cell survival of palate tissues. (A) Coronal palate sections of vehicle- or Olaparib-treated mouse embryos were labeled with BrdU (green) to detect proliferative cells, which were quantified. PS, palatal shelf. (B) TUNEL assay (green) and corresponding quantification of palate sections from vehicle- or Olaparib-treated mouse embryos. Arrows show TUNEL-positive signals. PS, palatal shelf. (C) Immunostaining for γ-H2AX (magenta, left panels), Caspase3 (Green, left panels) and γ-H2AX (magenta, right panels), p-Chk2 (green, right panels), and corresponding quantification in palate sections from vehicle- or Olaparib-treated mouse embryos at E13.5. The yellow boxes show the high-magnification image of γ-H2AX/Caspase3 and γ-H2AX/p-Chk2-positive cells. PS, palatal shelf. (D) Protein levels of γ-H2AX, Caspase3 and p53 were examined by Western blotting and quantified. (E) S9.6 intensity was examined using palatal cell lysates from vehicle- or Olaparib-treated mouse embryos at E13.5. Data in panels (A–D) are represented as mean ± SD, n = 3 in each group. Data in panel (E) are represented as mean ± SD, n = 10 in each group. *p < 0.05; **p < 0.01; N.S., not significant.