Fig. 1.
E(Pc) is required cell autonomously for GSC maintenance and germ cell differentiation. tub-Gal80ts, nos-Gal4 (Ctrl), and tub-Gal80ts, nos > E(Pc) shRNA [E(Pc) KD] testes at a, b permissive temperature (18 °C) and c–h restrictive temperature (29 °C), immunostained with Vasa to label germ cells, FasIII for hub (asterisk) and α-spectrin for spectrosome and fusome. i Quantification of GSCs number (average ± SD). Mann–Whitney U-test. n.s. nonsignificant, ****P < 0.0001. j–o GFP was used to identify Ctrl and E(Pc) mutant clones. j–k 2D after clonal induction (ACI), GFP-negative Ctrl GSC clone (yellow arrowhead in j), and E(Pc) mutant GSC clone (yellow arrowhead in k); GFP-negative Ctrl spermatogonial (SG) clone (cyan outline in j) and E(Pc) mutant SG clone (cyan outline in k). l, n 4D ACI, Ctrl GSC clone (yellow arrowhead), SG clone (cyan outline), and spermatocyte (SC) clone (magenta outline). m, o 4D ACI, E(Pc) mutant SG clone with bright DAPI intensity o has >16 germ cells within one cyst (cyan outline). No GSC clone or SC clone could be found for testes shown. p–r Percentage of testes with GSC clone p, SG q, SC r clones. N at the bottom represents the number of total testes from three independent clonal induction experiments. For each experiment, number of testes with at least one GFP-negative clone/total testes number was calculated and presented as % of testes with the corresponding type of clone. Asterisk: hub. Scale bar for f, g, h: 50 μm. Scale bar for other panels: 20 μm.