Fig. 5: dATR sustains tissue integrity in imaginal discs with reduced POLA2.

A-C Confocal images of ap > GFP (A), ap > GFP + dATR-i (B), and ap > GFP + dATM-i (C) discs stained with anti-pH2Av and anti-Dcp1. In the merge, DAPI, GFP, pH2Av, and Dcp1 are shown in blue, green, red, and grayscale, respectively. Scale bars, 100 µm. D-F Confocal images of ap > POLA2-i + GFP (GFP not shown) (D), ap > POLA2-i + dATR-i (E), and ap > POLA2-i + dATM-i (F) discs stained with anti-pH2Av and anti-Dcp1. In the merge, DAPI, pH2Av, and Dcp1 are shown in blue, red, and grayscale, respectively. Scale bars, 100 µm. G, H Quantification of apoptotic area per dorsal area (G) and of dorsal area (H) of the genotypes indicated in A–F (n = 20, 15, 23, 20, 37, 37) represented as a fold change normalized to control discs (ap > GFP). Note that the data points in G for ap > GFP and ap > POLA2-i + GFP are the same as in Fig. 3H and Fig. 6G, and the data points in H are the same as in Fig. 6H because these experiments were run in parallel. Data shown are mean ± SD. Statistical analysis was performed using a two-way ANOVA followed by a Tukey’s HSD test. Only the relevant one-to-one comparisons are shown. I, J Confocal images of the wing pouch of ap > POLA2-i + GFP (GFP not shown) (I) and ap > POLA2-i + dATR-i (J) discs labeled with anti-pH2Av and anti-Rad51. The DV boundary and outer border of the wing pouch are outlined in yellow. The Rad51 maxima panel exhibits the selection of Rad51 foci in the dorsal compartment. In the merge, DAPI, pH2Av, and Rad51 are shown in blue, red, and grayscale, respectively. Scale bars, 10 µm. K Quantification of the Rad51 foci per area in the dorsal compartment of the genotypes indicated in I and J (n = 19, 21). Note that the data points for ap > POLA2-i + GFP are the same as in Fig. 6K because these experiments were run in parallel. Data shown are mean ± SD. Statistical analysis was performed using a two-tailed unpaired t-test.