a-b, Confocal micrographs of isolated, male-derived spermatocytes (a) and budding spermatids (b) expressing GFP::3xFLAG::WAGO-3 and PEI-1::mTagRFP-T. Images were taken after a 30 minute treatment with 1,6-hexanediol. Hoechst33342 was used to stain DNA. Residual bodies are marked by a dashed circle. Images represent two biologically independent experiments. Scale bars: 4 μm. c, FRAP recovery curve of GFP::3xFLAG::WAGO-3 localizing to either P granules in L4 gonads or PEI granules in male-derived spermatids. Normalized data is presented as mean +/− SD and was fitted to a double exponential curve (n = 4 granules pooled from one independent experiment). d, Time sequence showing fluorescence recovery after photobleaching (FRAP) of GFP::3xFLAG::WAGO-3 localizing to either P granules in L4 gonads or PEI granules in male-derived spermatids. Residual bodies are marked by a dashed circle. Images represent two biologically independent experiments. Scale bars: 4 μm. e, Time sequence of GFP::3xFLAG::WAGO-3, taken from Extended Data Movie 1. Images are confocal maximum intensity projections of an isolated, male-derived spermatocyte. Images represent two biologically independent experiments. Scale bar: 4 μm. Source data are provided.