Figure 1. Centrosome age affects chromosome alignment.

(A) HeLa-eGFP-centrin1 (green) cell stained for cenexin (red, old centrosome marker) and DAPI (blue, DNA). One spindle pole contains the old centriole (brightest centrin1 signal and cenexin positive) and an accompanying daughter centriole (dim signal), which together form the old centrosome. The other spindle pole contains the young centriole (intermediate centrin1 signal), which is also accompanied by a daughter centriole and which together form the young centrosome. Scale bar in all panels = 5 μm. (B) Amounts of eGFP-centrin1 on the old, young and daughter centrioles in HeLa-eGFP-centrin1 cells determined from 3 independent experiments in 140 cells. (C) Untreated HeLa-eGFP-centrin1 cell (upper panel) and hTert RPE-eGFP-centrin1 cell treated with 10 ng/ml nocodazole (lower panel) stained for CENP-A (kinetochore marker) and DAPI. Yellow arrowheads indicate unaligned chromosomes; white arrowheads old centrosomes. (D and E) Proportion of unaligned chromosomes at old centrosomes in HeLa-eGFP-centrin1 cells (D), and in RPE1 cells stained for cenexin, RPE1-eGFP-centrin1 cells and HeLa-eGFP-centrin1 cells treated with 10 ng/ml nocodazole (E). For experiment and cell numbers, and p-values see Table 1. For results of individual experiments see Figure 1—source data 1. Error bars indicate s.e.m. * indicates p ≤ 0.05 in Binomial test compared to random distribution, *** indicates p ≤ 0.01 in Binomial test compared to random distribution.

DOI: http://dx.doi.org/10.7554/eLife.07909.003