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 Representative RPE-1 cells expressing CENP-A-GFP and centrin1-GFP (gray, centrioles circled) after indicated treatments, showing enlarged insets of polar (P) and aligned (A) kinetochores immunostained for pT232-Aurora B (red). b Ratio of pT232-Aurora B intensity normalized to CENP-A on polar vs. aligned kinetochores per cell under indicated treatments. 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. c Average pT232-Aurora B levels normalized to CENP-A on all kinetochore groups after indicated treatments. Dispersion measures as in ( b ). d Representative cells expressing CENP-A-GFP and centrin1-GFP (cyan, centrioles circled) immunostained for pT232-Aurora B (red) and α-tubulin (gray) after continuous CENP-E inhibition and acute Aurora A inhibition by MLN8054 and TCS7010, with enlarged insets of polar (P) and aligned (A) kinetochores. e – g Quantification of pT232-Aurora B intensity ratio on polar vs. aligned kinetochores e , number of polar chromosomes f , and spindle length g after indicated treatments. Dispersion measures as in ( b) . h RPE-1 cells expressing CENP-A-GFP and centrin-GFP (red, centrioles circled), immunostained for α-tubulin (gray), and imaged by super-resolution Airyscan microscopy showing enlarged insets of congressing (C) and aligned (A) kinetochores. Images are deconvolved projections of 2–5 z-planes. Numbers: b 23, 18, 14, 27 cells averaged from 491 kinetochore pairs, c 83, 71, 61, 57, 47, 41, 52, 79 kinetochore pairs from 91 cells, e 22, 19, 17, 18 cells averaged from 436 kinetochore pairs, f , g 39, 38, 42, 38 cells, all pooled from ≥2 independent biological replicates. Statistics: two-tailed ANOVA with post-hoc Tukey’s HSD test. Symbols indicate: n.s., p > 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001; inh., inhibited; chrom., chromosomes; MLN, MLN8237. Source data are provided as a Source Data file.
Article Snippet: Aurora A inhibitor TCS7010 (MedChemExpress, IC 50 value 3.4 nM) at a final concentration of 100 nM, was added 30 minutes before fixation.
Techniques: Expressing, Standard Deviation, Dispersion, Inhibition, Microscopy, Two Tailed Test