Fig. 4

PGC cyst-like division responds to nutritional status promptly. Confocal images of L3 ovaries of control (A), starved for 3 days (B), 8 h after refeeding (C), 16 h after refeeding (D), or 24 h after refeeding (E) with Vasa (green), α-Sepc/phalloidin (red), DNA (blue), and PH3 (white) staining. A A well-fed control L3 ovary shows PH3 + PGC cyst. B A L3 ovary after 3-day starvation does not contain proliferating PGC cyst. C A L3 ovary after 8 h refeeding does not contain proliferating PGC cyst. Starved L3 ovary shows PH3 + PGC cyst at 16 h (D) and 24 h (E) after refeeding. F Quantification of percentages of ovaries with PH3-positive PGCs from three to five pooled independent replicates for A–E using Fisher’s exact test. Confocal images of mock-treated (G) and rapamycin-treated L3 ovaries for 16 h (H) with Vasa (green), α-Spec/phalloidin (red), DNA (blue), and PH3 (white) staining. I Quantification of percentages of ovaries with PH3-postive PGCs from three pooled independent replicates for G and H using Fisher’s exact test. Confocal images of ovaries of wL4 (J, K) and 3-day females (L, M) with Vasa (green) and DNA (white) staining. Compared to control (J, L), the size of the ovaries reduces dramatically under long-term starvation (K, M). N–P Quantifications of ovariole number, egg number, and hatch rate of females from control and long-term starved larvae from three pooled independent replicates using non-parametric t-test. Data are represented as mean ± SEM. The number of samples (n) in each group is shown above the X-axis. ***p < 0.001.*p < 0.05. ns, not significant. The dash lines mark PH3 + PGC cysts. Scale bar in A indicates 50 µm (for A–E, G, H). Scale bar in J indicates 50 µm (for J–M)