Figure 6. p53A and p53B mutants have persistent germline DNA damage.
(A-D’) Images of ovarioles of indicated genotypes immunolabeled with anti-γ-H2Av (red) (A–D) to detect DNA breaks, and counterstained with the DNA dye DAPI (blue) (A’-D’). The ovarioles are oriented with the anterior germarium to left (square bracket). Scale bars are 25 µm. (A”-D”) Quantification of percent of ovarioles with γ-H2Av-positive nurse cells (blue squares) and oocyte (red ovals) for the indicated genotypes. Values are means of three biological replicates and >20 ovarioles, with error bars representing S.E.M. Values with low variance have very small error bars that are not visible in the graphs. See Figure 6—figure supplement 1 for higher magnification images of germaria, and Supplementary file 1 for p values.
Figure 6—source data 1. Quantification of persistent DNA breaks by stage for Figure 6.
elife-61389-fig6-data1.xlsx (186.8KB, xlsx)
Figure 6—figure supplement 1. Double labeling for oocyte marker Orb and γ-H2Av in stage 1.
(A-F’) Images of germarium and stage one labeled for γ-H2Av (red) and oocyte marker Orb (green) (A–F), and with DAPI DNA stain (blue) (A’-F’). (G) Quantification of percent stage one with γ-H2Av and Orb positive oocyte. (H) Quantification of percent stage one with a γ-H2Av -bright nurse cell adjacent to a Orb-positive oocyte. n = 20 ovarioles for each genotype.
Figure 6—figure supplement 1—source data 1. Quantification of DNA breaks in orb-lableled ovaries of Figure 6—figure supplement 1.
elife-61389-fig6-figsupp1-data1.xlsx (9.4KB, xlsx)
Figure 6—figure supplement 2. Quantification of γ-H2Av fluorescence intensity in oocytes (red) and nurse cells (blue) of stage one egg chambers.
Each dot represents a single oocyte or nurse cell. Horizontal lines represent mean and standard deviation. Asterisks represent adjusted p values of a two-way, pair-wise ANOVA computed through a Tukey test between p53A+B+ (wild type) and all other genotypes for oocyte (red asterisks) and nurse cells (blue asterisks). * p < 0.05; **p < 0.01; ***p < 0.01; ****p < 0.0001. Statistical analyses were performed using GraphPad Prism (version 9.1.2). See Supplementary file 2 for p values.
Figure 6—figure supplement 2—source data 1. Quantification of DNA breaks per cell for Figure 6—figure supplement 2.
elife-61389-fig6-figsupp2-data1.xlsx (19.7KB, xlsx)
Figure 6—figure supplement 3. DNA repair foci in p53 mutants depends on DNA meiotic breaks.
γ-H2Av labeling of wild type (A), mei-W681/ Df(2R)BSC782; p53A2.3 (B), and mei-681/Df(2R)BSC782; p53B41.5 (C).