GSCs and germaria with reduced CENP-C do not exhibit obvious defects in mitosis.
(A-D) 5 day old nanos-GAL4 and (A’-D’) CENP-C RNAi (germline tumour phenotype) stained with DAPI (cyan), VASA (grey) and H3S10P (red). * denotes cap cells. Scale bar = 10 μm. (E) Mitotic index (%) of H3S10P positive GSCs in nanos-GAL4 and CENP-C RNAi (n = 150 germaria). (F) Violin plot displaying the number of H3S10P positive cysts per germaria (n = 150 germaria). One positive hit was quantified as H3S10P positive GSC-CB pairs, 2-cell cysts (2cc), 4-cell cysts (4cc) or 8-cell cysts (8cc). ns = non-significant. (G-K) 1 day old nanos-GAL4 and (G’-K’) CENP-C RNAi stained with DAPI (cyan), histone H3 phosphorylated on threonine 3 (H3T3P) to mark prometaphase GCSs (blue), Spc105 (yellow) and 1B1 (red). 1 day old flies were analysed for this experiment in order to isolate prometaphase in actively dividing GSCs (circled). * denotes cap cells. Scale bar = 5 μm. (L-Q) nanos-GAL4 and (L’-Q’) CENP-C RNAi stained with Hoescht (cyan), 1B1 (red), CID (grey) and Cen3Giglio oligopaint FISH (yellow). White arrows indicate Cen3 that overlap with CID foci. GSCs are circled. (R) Quantitation of CID foci per GSC (grey bars) or Cen3 foci (overlapping with CID) per GSC (yellow bars) in nanos-GAL4 and CENP-C RNAi (n = 30 GSCs). ns = non-significant. Error bars = SEM.
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