CENP-C is required for CAL1 localisation in GSCs, but it is not required for CID localisation at later stages of development.

Immunofluorescent image of G₂/prophase GSCs (circled) in (A-C) nanos-GAL4 and (A’-C’) CENP-C RNAi stained for DAPI (cyan), 1B1 (red), CAL1 (yellow) and CENP-C (green). Centromeric CAL1 was identified as being colocalised with CENP-C. GSCs are circled. *denotes cap cells. Scale bar = 5 μm. (D) Quantitation of total centromeric CAL1 fluorescent intensity (integrated density) in nanos-GAL4 and CENP-C RNAi. ***p<0.001. Error bars = SEM. (E-H) bam-GAL4 and (E’-H’) bam-GAL4 driven CENP-C RNAi stained with DAPI (cyan), CENP-C (yellow) and 1B1 (red). Circle marks region where knockdown begins. (I-L) bam-GAL4 and (I’-L’) bam-GAL4 driven CENP-C RNAi stained with DAPI (cyan), H3S10P (red) to mark 8-cell cysts in mitosis (circled) and CENP-C (yellow). (M) Quantitation of CENP-C in each cell of 8-cell cysts of bam-GAL4 and CENP-C RNAi. **p<0.01. Error bars = SEM. (N-Q) bam-GAL4 and (N’-Q’) bam-GAL4 driven CENP-C RNAi stained with DAPI (cyan), H3S10P (red) to mark 8-cell cysts in mitosis (circled) and CID (yellow). (R) Quantitation of CID in each cell of 8-cell cysts of bam-GAL4 and CENP-C RNAi. ns = non-significant. Error bars = SEM. * denotes cap cells. Scale bar = 10 μm.

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