Fig 4. Centromere clustering is disrupted in Mcm5^A7 mutants.

a. Left: Representative images of centromere clustering, or lack thereof, in wild-type (WT) and Mcm5^A7 meiotic nuclei located in zygotene. Magenta: C(3)G, green: CID (centromere). In these images, WT nucleus contains 1 CID focus, and Mcm5^A7 contains 6 CID foci. Scale bar = 1 μm. Right: Quantification of CID foci in zygotene in WT (n = 24) and Mcm5^A7 (n = 16). ***p < 0.0001, unpaired T-test. Data are represented as mean ± SD. b. Left: Representative images of centromere clustering in WT and Mcm5^A7 early pachytene nuclei. Magenta: C(3)G, green: CID. In these images, WT nucleus contains 2 CID foci, and Mcm5^A7 contains 5 CID foci. Scale bar = 1 μm. Right: Quantification of early pachytene CID foci in WT (n = 65) and Mcm5^A7 (n = 94). ***p < 0.0001, unpaired T-test. Data are represented as mean ± SD. c. Left: Representative images of centromere clustering, or lack thereof, in mid-pachytene WT and Mcm5^A7 nuclei. Magenta: C(3)G, Green: CID. In these images, WT nucleus contains 1 CID focus, and Mcm5^A7 contains 4 CID foci. Scale bar = 1 μm. Right: Quantification of mid-pachytene CID foci in WT (n = 16) and Mcm5^A7 (n = 19). ***p < 0.0001, unpaired T-test. Data are represented as mean ± SD. d. Schematic representing relative location of 359-bp locus on Chromosome X (not drawn to scale). Top panel: Representative image of meiotic nucleus with 1 359-bp (green) focus (WT, Region 2A). Bottom panel: Representative image of meiotic nucleus with 2 359-bp (green) foci (Mcm5^A7, Region 2A). Right: Percentage of nuclei with paired 359-bp loci (one focus) or unpaired (two loci) in WT (n = 88) and Mcm5^A7 (n = 63) meiotic nuclei. ***p < 0.0001, as determined by two-tailed Fisher’s exact test. Contrast and brightness of all images were adjusted for clarity. Outlines of representative nuclei are drawn in white. CID foci not localized with C(3)G are from adjacent, non-meiotic cells. All DAPI-containing images are provided in S2 Fig.