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. Author manuscript; available in PMC: 2018 Jul 24.
Published in final edited form as: Dev Cell. 2017 Jul 24;42(2):130–142.e7. doi: 10.1016/j.devcel.2017.06.022

Figure 4. GCL controls PGC formation by inhibiting Torso activity at the posterior pole.

Figure 4

(A) Torso signaling was manipulated in gclΔ/+ control embryos by introducing tsl loss-of-function mutations. Number of PGCs in embryos from mothers of indicated genotype was counted and plotted. Bars represent the mean ± standard deviation. tsl−/− corresponds to tslA263V/Y279N. (n > 15, ****P < 0.0001, Mann-Whitney test)

(B) Torso expression was manipulated by combining gclΔ/Δ mutation with transgenic-based torso knockdown (RNAi) with three individual shRNA lines. The germline-specific driver maternal tubulinGAL4∷VP16 was used to express shRNA. Non-targeting RNAi against mCherry in gclΔ/Δ embryos was used as a control. Number of PGCs in embryos from mothers of indicated genotype was counted and plotted. Bars represent the mean ± standard deviation. (n = 10, ****P < 0.0001, Mann-Whitney test)

(C) Torso signaling was manipulated in gclΔ/Δ embryos by introducing tsl loss-of-function mutations. Number of PGCs in embryos from mothers of indicated genotype was counted and plotted. Bars represent the mean ± standard deviation. Tsl−/− corresponds to tslA263V/Y279N. (n > 20, ****P < 0.0001, Mann-Whitney test)

(D) Representative embryos of indicated maternal genotype. Anti-Vasa (green) immunostaining marks PGCs in the embryos, and F-actin labeled by phalloidin (magenta) outlines both germ and somatic cells. Scale bar = 20μm.

(E) Fixed embryos from mothers of indicated genotype were immunostained with anti-Vasa (green) and anti-RNA PolymeraseII phosphoSer2 (RNAPII pSer2) (red). DNA (blue). To image a uniform layer of PGCs, posterior portion of the embryos was cut, turned, and mounted. Dotted line outlines Vasa-expressing PGCs. Scale bar = 20μm.

(F) Fixed embryos from mothers of indicated genotype were immunostained with anti-Vasa (green) and anti-Torso (red). DNA (blue). Dotted line outlines Vasa-expressing PGCs. In gclΔ/Δ, tsl−/− embryo, but not gclΔ/+, tsl−/−, these cells express Torso at the cell membrane. Scale bar = 20μm.

See also Figures S4 and S5.