Figure 6.

Anterior-central grk mRNA localization in saturn is due to joint inactivation of klc and piRNA pathway. (A−E) Sagittal views as well as transverse projections of a series of z-stack confocal images of endogenous grk mRNA visualized by in situ hybridization in wildtype (A), saturn (B), klc^sat (C), mael^sat (D), and armi^3R13 klc^8ex94 double mutant (E) stage 8−10 egg-chambers. Nuclear DNA in the saturn oocyte is marked by arrow (B′), showing that the majority of grk mRNA is not associated to the nucleus. Approximately one-half the grk mRNA (arrowhead, C) is internalized in klc^sat germline clones. grk mRNA is also seen in the opposite side of the anterior periphery (arrowhead, D) in mael^sat germline clones. (F, G) Anti-Dhc staining in wild-type (F) and in saturn (G) stage 8−9 egg chambers. Dhc is excluded from the nucleus, thus showing the mispositioned nucleus. (H, I, J) grk*mCherry localization in stage 6 oocytes. (H) wild-type (wt), (I) klc^sat, and (J) klc^8ex94 showing internally localized grk mRNA in klc mutants. (K, L) Anti-Grk staining in stage 8−9 egg-chambers. (K) wildtype (wt). (L) klc^sat, showing that Grk translation is not disrupted in klc mutant. (M, N) Localization (arrows) of grk*mCherry (grk, M and N) and the dynein cofactor, BicD (M′ and N′) indicates that dynein-dependent grk mRNA transport is not affected in klc^sat germline clone. BicD and grk mRNA are excluded from the oocyte nucleus, showing a posteriorly mislocalized nucleus before its migration to the dorsoanterior corner. (O) Frequencies of abnormal or absent dorsal appendages in mature eggs of indicated genotypes. Scale bars = 5 μm in (M) and 20 μm in the rest.