Journal: Nature Communications

Article Title: Kinetochore-centrosome feedback linking CENP-E and Aurora kinases controls chromosome congression

doi: 10.1038/s41467-025-64804-1

Figure Lengend Snippet: a Schematic of the protocol used to modulate CENP-E activity in cells with variable centriole numbers per spindle pole, generated by continuous Plk4 inhibition. Numbers on the left denote number of centrioles per spindle pole. b Diagram showing progressive centriole depletion after continuous Plk4 inhibition by 300 nM centrinone. c Representative images of live RPE-1 cells expressing CENP-A-GFP and centrin1-GFP (color-coded by depth, color bar) with varying centriole counts (white circles), 5 minutes after end of prometaphase spindle elongation, under indicated treatments. d , e Quantification of polar chromosome number d , and their residence time at spindle poles e , after prometaphase spindle elongation, comparing cells with different centriole numbers. Category “1” includes centriolar poles from both 1:1 and 1:0 spindles. Colored points represent individual cells; black lines show the mean, with light and dark gray areas marking 95% confidence intervals for the mean and standard deviation, respectively. Number of cells per condition: 14, 15, 22, 16, 14, 18, 16, 16, 12, 12, 16, 16 ( d , e ), each pooled from ≥3 independent biological replicates. f Number of polar chromosomes after treatments in cells with varying centriole numbers, fixed prior to measurement. Pooled data from ≥3 independent biological replicates. Number of kinetochores and cells is given in the figure. Dispersion measures as in ( d , e ). g STED microscopy images of RPE-1 cells immunostained for α-tubulin (gray), expressing CENP-A-GFP and centrin1-GFP (color-coded by depth). All images are maximum projections. Statistics: two-tailed ANOVA with post-hoc Tukey’s HSD test. Symbols indicate: n.s., p > 0.05; *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001; inh., inhibited; depl., depleted; siRNA, small interfering RNA; KC, kinetochore; ctrl., control. Source data are provided as a Source Data file.

Article Snippet: Plk4 inhibitor Centrinone (MedChemExpress, K i value 0.16 nM) at a final concentration of 300 nM, was added 1-5 days before the imaging as noted in the manuscript.

Techniques: Activity Assay, Generated, Inhibition, Expressing, Standard Deviation, Dispersion, Microscopy, Two Tailed Test, Small Interfering RNA, Control