Figure 5. Translocation of Piwi–pre‐piRISC from Yb bodies requires the RNA‐binding activity of Armi.

1. Armi‐Flag (Armi‐F) WT and the N756A mutant were expressed in OSCs where endogenous Armi and Zuc had been depleted by RNAi (Zuc KD + Armi KD). Immunoprecipitation and subsequent Western blotting show that Armi‐F WT bound with Piwi and Gasz. The Armi N756A mutant bound with Piwi strongly but only weakly with Gasz. Luciferase‐Flag (Luc‐F) was used as a negative control.
2. Armi‐Flag (Armi‐F) WT and the N756A mutant were expressed in OSCs where endogenous Armi and Zuc had been depleted by RNAi (Zuc KD + Armi KD). Armi‐F WT (green) localized onto mitochondria (red), whereas Armi‐F N756A mutant (green) localized to Yb bodies (see also Fig EV5D). The scale bar represents 5 μm. DAPI (blue) shows the nuclei.
3. In vitro pull‐down assays show that both Armi‐Flag (Armi‐F) WT and N756A mutant directly bind with GST‐GaszΔC113 but not with GST. Armi‐F WT and mutant were immunopurified from Schneider 2 (S2) cells under harsh conditions. GST and GST‐GaszΔC113 were visualized by CBB staining, while Armi‐F WT and N756A mutant were detected by Western blotting using anti‐Flag antibodies.