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. 2022 Jan 13;11:e61389. doi: 10.7554/eLife.61389

Figure 7. p53A and p53B have overlapping and distinct functions in germline genome integrity and the meiotic pachytene checkpoint.

(A-E’) Drosophila ovarioles of indicated genotypes were immunolabeled for γ-H2Av (red A-E) to detect DNA breaks and counterstained with the DNA dye DAPI (blue A’-E’). The ovarioles are shown with the anterior germarium to left (square bracket). Scale bars are 25 µm. (A”-E”) Quantification of percent of ovarioles with γ-H2Av-positive nurse cells (blue squares) and oocyte (red ovals) at different stages. Values are means of five biological replicates and >20 ovarioles with error bars representing S.E.M. Those values that had low variance have very small error bars that are not visible in the graphs. See Figure 6—figure supplement 1 for higher mag images of germaria, and Supplementary file 1 for p values. (F–K) p53A is required for activation of the pachytene checkpoint. (F–J) Oocyte nuclei from stage 3 to 4 egg chambers labeled with antibodies against synaptonemal protein C(3)G (red) and DNA dye DAPI (blue). (F) Wild type with spherical compact karyosome. (G) okraRU / okraAA with diffuse chromatin indicating activation of the pachytene checkpoint. (H) okraRU / okraAA; p535A-1-4 (A-B-) null with compact spherical karyosome. (I) okraRU / okraAA; p53A2.3 (A-B+) p53A mutant with spherical karyosome. (J) okraRU / okraAA; p53B41.5 (A+B-) with elliptical nucleus. Scale bars are 3 µm. (K) Quantification of karyosome formation. Data are means based on two biological replicates with ~30 nuclei per strain per replicate, with error bars representing S.E.M. * p < 0.05, ** p < 0.01, n.s. = not significant by unpaired Student’s t test.

Figure 7—source data 1. Quantification of persistent DNA breaks by stage for Figure 7.
elife-61389-fig7-data1.xlsx (186.8KB, xlsx)
Figure 7—source data 2. Quantification of karysome phenotype for Figure 7K.

Figure 7.

Figure 7—figure supplement 1. Images of eggshell phenotype classes produced by p53 and okr single or double mutant mothers.

Figure 7—figure supplement 1.

p53A+B+ = wild type. okr; p53A+B+ = okra single mutant. Class I- wild type; Class II: ventralized; Class III: short or missing dorsal appendages. Class IV: Small collapsed eggs with thin eggshells. See Figure 7—figure supplement 2 for quantification.
Figure 7—figure supplement 2. Quantification of eggshell phenotype classes produced by p53 and okr single or double mutant mothers.

Figure 7—figure supplement 2.

p53A+B+ = wild type. okr; p53A+B+ = okra single mutant. Class I- wild type; Class II: ventralized; Class III: short or missing dorsal appendages. Class IV: Small collapsed eggs with thin eggshells.
Figure 7—figure supplement 2—source data 1. Quantification of egg phenotypes for Figure 7—figure supplement 2.
Figure 7—figure supplement 3. p53 mutant mothers have reduced fertility.

Figure 7—figure supplement 3.

Hatch rates of eggs from p53 and okr single or double mutant mothers. The hatch rate for all okr single and double mutants was zero (complete female sterility). p53A+B+ = wild type. okr; p53A+B+ = okra single mutant. Shown are mean and standard deviation based on four to five biological replicates of 30–50 eggs per genotype. *p < 0.05, **p < 0.01 versus wild type by unpaired student’s t test.
Figure 7—figure supplement 3—source data 1. Quantification of hatch rates for Figure 7—figure supplement 3.