Figure 2.
Centrosome-age-dependent asymmetric spindles’ asymmetric daughter cell size. (A) Image of a live 2:2 RPE1 GFP-centrin1 mScarlet-cenexin metaphase cell. (B) Quantification of the SAI versus centrosome age in live cells. On average, the mean SAI is 2.7 ± 5.5%, n = 28 cells, P = 0.0145. At t = −1 min before anaphase onset, the mean SAI is 3.8 ± 5.6%, n = 28 cells, P = 0.0012 in one-sample Wilcoxon tests. (C) Immunofluorescence images of 2:2, 1:1, and 0:0 RPE1 GFP-centrin1 late telophase cells. Cells are stained with DAPI (blue), GFP-centrin1 (green), cenexin (yellow) antibody and the cell mask (magenta). (D) Quantification of the daughter cell area asymmetry index. For 2:2 cells = 2 ± 5.3%, n = 65 cells, P = 0.0025; for 1:1 cells = 4.7 ± 7.5%, n = 58 cells, P <0.0001, and for 0:0 cells = 0.25 ± 10.5%, n = 70 cells, P = 0.8370 in one-sample t tests. (E) Immunofluorescence images of 2:2 and 1:1 RPE1 GFP-Centrin1 metaphase cells stained with DAPI (blue), GFP-centrin1 (green), cenexin antibody (yellow) and the cell mask (magenta). (F) Quantification of the cortex-centrosome distances in fixed and live cells. Mean distances of 6.8 ± 1.2 μm and 7.1 ± 1.5 μm for the old and young centrosomes, respectively (44 cells, P = 0.2135, ns in paired t test) in fixed cells and means of 6.7 ± 1.7 μm and 7.3 ± 2 μm for the old and young centrosomes, respectively (28 cells, P = 0.4491, ns in paired t test) in live cells 1 min before anaphase onset. (G) Immunofluorescence image of a 1:1 centriolin−/− GFP-centrin1 RPE1 late telophase cell stained with DAPI (blue), GFP-centrin1 (green), cenexin antibody (yellow) and the cell mask (magenta). (H) Quantification of the daughter cell area asymmetry index of 1:1 centriolin−/− GFP-centrin1 RPE1 cells = −0.4 ± 5.8%, n = 54 cells, P = 0.6246, in one-sample t test. All scale bars = 10 μm. ns = not significant; P ≤ 0.05 = *; P ≤ 0.01 = **; P ≤ 0.0001 = ****.

Centrosome-age-dependent asymmetric spindles’ asymmetric daughter cell size. (A) Image of a live 2:2 RPE1 GFP-centrin1 mScarlet-cenexin metaphase cell. (B) Quantification of the SAI versus centrosome age in live cells. On average, the mean SAI is 2.7 ± 5.5%, n = 28 cells, P = 0.0145. At t = −1 min before anaphase onset, the mean SAI is 3.8 ± 5.6%, n = 28 cells, P = 0.0012 in one-sample Wilcoxon tests. (C) Immunofluorescence images of 2:2, 1:1, and 0:0 RPE1 GFP-centrin1 late telophase cells. Cells are stained with DAPI (blue), GFP-centrin1 (green), cenexin (yellow) antibody and the cell mask (magenta). (D) Quantification of the daughter cell area asymmetry index. For 2:2 cells = 2 ± 5.3%, n = 65 cells, P = 0.0025; for 1:1 cells = 4.7 ± 7.5%, n = 58 cells, P <0.0001, and for 0:0 cells = 0.25 ± 10.5%, n = 70 cells, P = 0.8370 in one-sample t tests. (E) Immunofluorescence images of 2:2 and 1:1 RPE1 GFP-Centrin1 metaphase cells stained with DAPI (blue), GFP-centrin1 (green), cenexin antibody (yellow) and the cell mask (magenta). (F) Quantification of the cortex-centrosome distances in fixed and live cells. Mean distances of 6.8 ± 1.2 μm and 7.1 ± 1.5 μm for the old and young centrosomes, respectively (44 cells, P = 0.2135, ns in paired t test) in fixed cells and means of 6.7 ± 1.7 μm and 7.3 ± 2 μm for the old and young centrosomes, respectively (28 cells, P = 0.4491, ns in paired t test) in live cells 1 min before anaphase onset. (G) Immunofluorescence image of a 1:1 centriolin−/− GFP-centrin1 RPE1 late telophase cell stained with DAPI (blue), GFP-centrin1 (green), cenexin antibody (yellow) and the cell mask (magenta). (H) Quantification of the daughter cell area asymmetry index of 1:1 centriolin−/− GFP-centrin1 RPE1 cells = −0.4 ± 5.8%, n = 54 cells, P = 0.6246, in one-sample t test. All scale bars = 10 μm. ns = not significant; P ≤ 0.05 = *; P ≤ 0.01 = **; P ≤ 0.0001 = ****.

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