Fig. 5.

Set1 regulates key JAK-STAT and BMP signaling pathway components. (A) Representative images of nos>set1 RNAi, mad¹²/+ and nos-Gal4/mad¹² (on the 2nd chromosome, abbreviated as mad¹²/+); nos>set1 RNAi testes at 5 days post-eclosion, immunostained for pMad (green), Fas III (red) for the hub region, 1B1 (red) for spectrosome and fusome, and Tj (magenta) for the CySC lineage cells. (B) Representative images of nos>set1 RNAi, stat92E⁰⁶³⁴⁶/+ and UAS-set1 RNAi/stat92E⁰⁶³⁴⁶ (on the 3rd chromosome, abbreviated as stat92E⁰⁶³⁴⁶/+); nos>set1 RNAi testes at 5 days post-eclosion, immunostained for Stat92E (green), Fas III (red) for the hub region, 1B1 (red) for spectrosome and fusome, and Tj (magenta) for the CySC lineage cells. In B, yellow dotted outline delineates the hub, white dashed outline the GSCs and CySCs region, and cyan dashed outline the spermatogonial region. Scale bars: 20 μm. (C) Quantification of the percentage of testes with representative Stat92E antibody expression: normal or ectopic with prolonged expression patterns present in stat92E⁰⁶³⁴⁶/+ versus nos>set1 RNAi versus nos>set1 RNAi; stat92E⁰⁶³⁴⁶/+ testes at 5 days post-eclosion. *P<0.05, χ² test. (D) Quantification of the percentage of testes with representative pMad antibody expression: normal or ectopic with prolonged expression patterns present in mad¹²/+ versus nos>set1 RNAi versus mad¹²/+; nos>set1 RNAi testes at 5 days post-eclosion. n.s., not significant, χ² test. (E) Quantification of the percentage of testes with the representative germline phenotypes, germ cell loss and early-germline overpopulation, using the same criteria as defined in Fig. 1D, present in mad¹²/+ (n=56), tkv⁴/+ (n=45), stat92E⁰⁶³⁴⁶/+ (n=40), nos-Gal4>set1 RNAi (n=84), mad¹²/+; nos-Gal4>set1 RNAi (n=43), tkv⁴/+; nos-Gal4>set1 RNAi (n=39) and stat92E⁰⁶³⁴⁶/+; nos-Gal4>set1 RNAi (n=38) testes at 5 days post-eclosion. *P<0.05, ****P<10⁻⁴, χ² test. (F) Quantification of GSC number for mad¹²/+ (n=56), tkv⁴/+ (n=45), stat92E⁰⁶³⁴⁶/+ (n=40), nos-Gal4>set1 RNAi (n=84), mad¹²/+; nos-Gal4>set1 RNAi (n=43), tkv⁴/+; nos-Gal4>set1 RNAi (n=39) and stat92E⁰⁶³⁴⁶/+; nos-Gal4>set1 RNAi (n=38) testes at 5 days post-eclosion. Individual data points and mean values are shown. See also Table S1. Error bars represent s.e.m. *P<0.05, ***P<10⁻³, ****P<10⁻⁴; n.s., not significant; one-way ANOVA and Dunnett's T3 multiple comparison test was used to compare multiple individual datasets with each other that had one independent variable.