Fig. 3: Regulation of apoptosis in imaginal discs with POLA2 knockdown.

A, B Confocal images of ap > GFP + POLA2-i wing imaginal discs stained with anti-Dcp1. Transgene expression was induced for either 2 days (A) or 4 days (B). The basal side of the disc is displayed in A. In the merge, DAPI, GFP, and Dcp1 are shown in blue, green, and grayscale, respectively. Scale bars, 100 µm. C Confocal image of an ap > POLA2-i disc expressing hid-GFP and stained with anti-pH2Av. The dorsal compartment is outlined in yellow. In the merge, DAPI, hid-GFP, and pH2Av are shown in blue, green, and red, respectively. Scale bar, 100 µm. D Confocal image of an ap > POLA2-i disc expressing TRE-RFP and stained with anti-pH2Av. The dorsal compartment is outlined in yellow. In the merge, DAPI, pH2Av, and TRE-RFP are shown in blue, green, and red, respectively. Scale bar, 100 µm. E-G Confocal images of ap > POLA2-i + GFP (GFP not shown) (E), ap > POLA2-i + p53-i (F), and ap > POLA2-i+bsk-DN (G) 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. H Quantification of apoptotic area per dorsal area represented as a fold change normalized to control discs (ap > GFP). The analyzed genotypes were ap > GFP (n = 20), ap > GFP + p53-i (n = 20), ap > GFP+bsk-DN (n = 21), and the three genotypes indicated in E–G (n = 20, 25, 37). Note that the data points for ap > GFP and ap > POLA2-i + GFP are the same as in Fig. 5G and Fig. 6G 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.