The phage gene wmk is a candidate for male killing by a bacterial endosymbiont
Fig 5
Transgenic expression of wmk causes DNA damage in association with H4K16ac.
Images and data are from embryos 4–5 h AED expressing a transgene under the arm driver. (A) DAPI DNA stain of male and female embryos, side-by-side, expressing wmk. Sexes determined by H4K16ac antibody. (B) pH2Av antibody staining of the same embryos as (A). The male has distinct punctae or foci, while the female does not. All embryos exhibit either a low level of autofluorescence at the same wavelength as the secondary antibody (Alexa 488) visible in both embryos or there is background staining. (C) H4K16ac antibody staining of the same embryos as (A). Distinct punctae are only visible in males, while females can exhibit low levels of staining. (D) DAPI DNA stain of control gene male. Sex determined by H4K16ac antibody. (E) pH2Av antibody staining of the same embryo as (D), with no distinct punctae and only autofluorescence or background staining visible. (F) H4K16ac antibody staining of the same embryos as (D). (G) Graph of the number of pH2Av punctae visible in each embryo. N = 25 embryos per genotype. Statistics are based on a Kruskal-Wallis one-way ANOVA followed by Dunn’s correction. (H) Graph of the number of H4K16ac punctae visible in each of the same embryos as measured in (G). Statistics are based on a Mann-Whitney U test comparing the two male categories. (I) Number of cases where pH2Av punctae directly overlapped with H4K16ac punctae in the same embryos as (G) and (H). Statistics are based on a Kruskal-Wallis one-way ANOVA followed by Dunn’s correction. (J) Graph of the total number of chromatin bridges and the total number of bridges with overlapping H4K16ac and pH2Av punctae in each of the three genotypes measured in (G-I). All images were taken at 20X zoom. This experiment has been performed once. *p<0.05, ***p<0.001, ****p<0.0001.