Figure 3.

Heph and Hrb27C are necessary for timely cyst divisions and GSC proliferation, respectively. (A-D) Mock mosaic germarium from a female raised on a nutrient-deficient diet (control, A), hephe2 (B), sqdix50 (C), and Hrb27CrF680 (D) mosaic germaria stained with anti-GFP (green; wild-type cells), anti-cleaved Caspase (magenta, dying cysts), anti-Hts+anti-LamC (red; fusomes, follicle cell membranes, and cap cell nuclear envelopes). Since germaria from wild-type flies raised on nutrient-rich media rarely have dying cysts, a nutrient-deficient diet was used for one set of controls to ensure staining conditions for the cleaved-Caspase antibody were sufficient. (E) Percentage of total control (GFP+) or mutant (GFP-) cystoblasts/cysts in hnRNP mosaic germaria. Numbers in bars represent number of germline-mosaic germaria analyzed. Cyst division ratios are listed above bars (number of cysts scored in parentheses). (F-I) Mock mosaic (F), sqdix50 (G), hephe2 (H), and Hrb27CrF680 (I) mosaic germaria stained with anti-GFP (green; wild-type cells), EdU (white; cells in S phase), anti-Hts+anti-LamC (red). (J) Percentage of GFP-negative GSCs that are positive for EdU in mosaic germaria eight days after heat shock. Numbers in bars represent the number of GFP-negative GSCs analyzed. **p < 0.001, Chi-squares test. GSCs are outlined in white and cystoblasts/cysts are outlined in yellow (wild-type = solid line; mutant = dashed line); asterisks indicate cap cells. A caspase-positive wild-type cyst is outlined in yellow in A. Scale bars = 5 μm