Wolbachia endosymbionts manipulate the self-renewal and differentiation of germline stem cells to reinforce fertility of their fruit fly host

The alphaproteobacterium Wolbachia pipientis infects arthropod and nematode species worldwide, making it a key target for host biological control. Wolbachia-driven host reproductive manipulations, such as cytoplasmic incompatibility (CI), are credited for catapulting these intracellular bacteria to high frequencies in host populations. Positive, perhaps mutualistic, reproductive manipulations also increase infection frequencies, but are not well understood. Here, we identify molecular and cellular mechanisms by which Wolbachia influences the molecularly distinct processes of germline stem cell (GSC) self renewal and differentiation. We demonstrate that wMel infection rescues the fertility of flies lacking the translational regulator mei-P26, and is sufficient to sustain infertile homozygous mei-P26-knockdown stocks indefinitely. Cytology revealed that wMel mitigates the impact of mei-P26 loss through restoring proper pMad, Bam, Sxl, and Orb expression. In Oregon R files with wild-type f..., Candidate gene selection
Leveraging what is known about wMel’s abilities to rescue essential maintenance and differentiation genes with empirical data on protein-protein and protein-mRNA interactions among these genes (esyN3 networks in fig S1A-D and Fig 8; references in table S1) and data on interactions between host genes and wMel titers 4, we identified the essential germline translational regulator meiotic P26 (mei-P26) as a potential target of of bacterial influence over germline stem cell (GSC) maintenance and differentiation pathways (fig S1A,B). Loss of mei-P26 in uninfected flies produces mild to severe fertility defects through inhibiting GSC maintenance, meiosis, and the switch from germline cyst proliferation and differentiation. 
Drosophila stocks and genetic crosses 
Flies were maintained on white food prepared according to the Bloomington Drosophila Stock Center (BDSC) Cornmeal Food recipe (aka. “white food”, see https://bdsc.indiana.edu/information/recipes/bloomfood.html..., Fiji (ImageJ), This data repository contains the data for the fertility plots, confocal images, and fluorescence plots in the manuscript titled \"Wolbachia endosymbionts manipulate the self-renewal and differentiation of germline stem cells to reinforce fertility of their fruit fly host\".

There are tsv files for egg lay, egg hatch, and offspring counts, for all genotype and infection combinations reported in the paper, as indicated in the file names. These data are plotted in Figure 1 and 6 and supplementary figures S4, S8, S9.

The raw Leica sample image files used to generate the micrographs and fluorescence intensity plots in Figures 1-6 and supplementary figures S5 and S7 are included (.lif files).

There are also two tsv files for the fluorescence intensities from Bam and Sxl immunocytochemistry data points plotted in Figures 3 and 4 and supplementary figure S7.

The R scripts written and used to plot these data and analyze their significance are also included.

Leica .lif file naming (e.g., bam-...