Fig. 5. Vas-eIF5B interaction is important for the establishment of dorsoventral polarity in the egg, and for grk regulation. Dark-field photographs of dorsal appendage phenotypes (A-F). Eighty-two percent of vas^PH165;P{vas⁺} eggs have two dorsal appendages (A). Dorsal appendages in vas^PH165;P{vas⁶¹⁷} eggs reveal a range of phenotypes: 22% have two dorsal appendages (B), 67% exhibit one semi-fused or fully fused dorsal appendage (C-E) and 11% have no dorsal appendages (F). (G-L) grk RNA and Grk protein were visualized in the early stages of oogenesis, through in situ hybridization and immunostaining. (J-L) Grk protein is shown in red; DAPI staining of DNA is shown in blue. (G,J) In vas^PH165;P{vas⁺} ovaries, both RNA and protein are enriched in the developing oocyte. (H-L) In both vas^PH165 and vas^PH165;P{vas⁶¹⁷} ovaries, grk RNA is enriched in the oocyte (H,I); however, Grk protein is barely detectable (K,L). Grk protein was examined in single confocal sections using the same laser settings for each genotype.