Extended Data Fig. 8: CENP-A and NDC80 localization on the dicentric ROB chromosome in GM03417.
From: The formation and propagation of human Robertsonian chromosomes
A-B. Representative SIM images of ROB chromosome (A) and normal copies of chromosomes 14 and 21 (B) labeled by immuno-FISH with centromeric satellite probes for Cen 14/22 (orange), Cen 13/21 (green), and anti-CENP-A antibody (white). DNA was counterstained with DAPI. Plots below show averaged line intensity profiles along the individual kinetochore regions of sister chromatids normalized to the maximum intensity of each channel for 20 ROBs (A) and 11 normal copies of chr.14 and chr.21 (B). Error bars denote standard deviations. C. Example SIM images of four individual ROB chromosomes labeled by immuno-FISH with a probe for Cen 13/21 (green), and antibodies against CENP-A (white) and NDC80 (magenta). Magnified insets show centromeric regions labeled with each antibody individually. Line scans (1 and 2) along kinetochores of sister chromatids show normalized fluorescence intensity profiles for CENP-A (blue) and NDC80 (magenta), illustrating variations between kinetochores. D. SIM images and corresponding line scans of representative normal copies of acrocentric chromosomes 13 and 21 labeled and analyzed as in (C). Scale bar in all images is 1 µm.