Figure 3. Pum and Its Cofactor, Nos, Regulate Pgc Translation in the GSCs and Early-Differentiating Cysts.

(A) A germarium of a pgcGFP ovary stained with 1B1 (red), Vasa (blue), and GFP (green) shows expression of GFP only in the pre-CB (dashed circle).

(B) A germarium of a pgcGFP; pum^ET1/FC8 ovary stained with 1B1 (red), Vasa (blue), and GFP (green) shows aberrant GFP expression from GSCs to the 8-cell cyst (100% from GSC to 4-cell cyst, 32% in 8-cell cyst, n = 25) (dashed outline).

(C) A germarium of a pgcGFP; nos^RC/BN ovary stained with 1B1 (red), Vasa (blue), and GFP (green) shows aberrant GFP expression from GSCs to the 4-cell cyst (100% from GSCs to 2-cell cyst, 13% in 4-cell cyst, n = 25) (dashed outline).

(D) A germarium of a pgcGFP; twin^RY3/RY5 ovary stained with 1B1 (red), Vasa (blue), and GFP (green) shows aberrant GFP expression from GSCs to the 8-cell cyst (100% from GSC to 4-cell cyst, 40% in 8-cell cyst, n = 25) (dashed outline). The GFP channel is shown in A1–D1.

(E) A developmental profile of GFP expression when Pum, Nos, and Twin are depleted in the germline.

(F) PAT assay of pgc poly(A)-tail length in GSC tumors and in GSC tumors lacking Pum and Nos.

(G, I, and K) Germaria of pum^ET1/FC8 (G), nos ^RC/BN (I), and twin ^RY3/RY5 (K) mutants stained with 1B1 (red) and, Vasa (green).

(H, J, and L) Germaria of pgc; pum^ET1/FC8 (H) pgc; nos^RC/BN (J) and pgc; twin^RY3/RY5 (L) double mutants stained with 1B1 (red) and Vasa (green).

(M) A graphical representation of the rescue experiment (n = 40). A population proportion z-test was performed. ** and *** indicate a p value < 0.005 and < 0.0005, respectively.

Scale bars, 10 μm. See also Figures S3 and S4.