GFP::Spn-E localization is regulated by Qin and PIWI proteins. (A) Confocal micrographs of GFP::Spn-E in stage 3 egg chambers of wild type and mutants, as indicated. (Left) One egg chamber section. Bar, 5 µm. (Right) Magnified images of the perinuclear and nuclear regions of one nurse cell. Bar, 1 µm. (B) Relative sizes of nuage granules positive for GFP::Spn-E as measured by pixel number of individual granules. Error bars are SEM, and p-value is the result of a two-tailed t test comparing the difference in sizes between wild type (WT) and qin. n = 22 replicates. (C) Quantification of GFP::Spn-E fluorescence intensity in defined regions of nurse cells at stage 3 of oogenesis. Shown are mean intensities detected in peripheral cytoplasm and in individual nuage granules. Error bars are SEM, and the p-values are results of two-tailed t tests comparing the difference in intensities between wild type and qin. n = 23 replicates. (D) Localization of GFP::Spn-E and Ago3 proteins in a stage 3 egg chamber. Bar, 5 µm. (E) Confocal micrographs of GFP::Spn-E in stage 4 egg chambers of mutants, as indicated. (Left) Section through one egg chamber. Bar, 5 µm. (Right) Magnified images of the perinuclear and nuclear regions of one nurse cell. Bar, 1 µm. (F) GFP::Spn-E localization in wild-type and aub ago3 qin triple mutant stage 4 egg chambers. Bar, 5 µm.