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Fig. S4
Supporting information for Fig 3.(A) Quantification of mitochondrial enrichment in zygotene bouquet TEM image overlapping high magnification images of the entire cytoplasm 360° around the nucleus of the cell in the top panel are shown in the lower panels. Image numbers (bottom) correspond to the numbered regions (top). SC-NE contact points (yellow arrowheads) mark the presumptive telomere cluster. Mitochondria (examples are marked by yellow arrows) were counted in regions adjacent to SC-NE contact points versus regions that are not. In this example, frames 4–6 span the presumptive telomere cluster. Mitochondria are mostly found in the cytoplasm apposing this region versus that of frames 1–3 and 7–8. Pooled data are plotted in Fig 4A (n = 7 oocytes). Zygotene oocytes were identified by their size (measurements are shown in A), the detection of SC, and the typical peripheral nucleolus. Nuc, nucleus; Cyto, cytoplasm; nucl, nucleolus. Green arrowhead, NE; red arrowhead, cell membrane. Red arrowheads in the entire cell (top) indicate to the same regions in the corresponding smaller higher power images. (B) Zygotene nest analysis for DCLK-GFP, as shown in Fig 3B for DiOC6 and Buc. DCLK appears randomly radially distributed and shows no enrichment in the centrosome cytoplasm of either oogonia or zygotene oocytes (n = 5 ovaries Zygotene, n = 26 oocytes in 4 nests, Oogonia, n = 6 oocytes in 2 nests). Data in S1 Data. (C) mAb414 detects the NE and colocalizes with perinuclear granules in zebrafish oocytes. The mAb414 perinuclear spherules signal (distinct from the fine line of the NE) colocalizes with the piRNA-specific proteins Vasa (n = 8 ovaries), GasZ (n = 7 ovaries), Zili, (n = 4 ovaries), and Ziwi (n = 4 ovaries), here shown in oogonia. GasZ oocyte shown is the same cell shown in Fig 3C, but also showing the different channels separately. (D) GasZ is a Bb resident protein during mid-to-late diplotene stages. GasZ colocalizes with dazl mRNA (costained with DAPI, blue) in the mature Bb (arrowheads; n = 8 ovaries).