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. Author manuscript; available in PMC: 2016 Jun 5.
Published in final edited form as: Science. 2015 Apr 30;348(6239):1155–1160. doi: 10.1126/science.aaa5111

Fig. 3.

Fig. 3

Centrosome loss triggers a p53-dependent arrest in normal cells. (A) Centro- some number distribution (left; data are means T SD; N = 3) and proliferation (right, data are means T SEM; N = 3) of RPE1 cells after centrinone addition. (B) (left) Centro- some number distribution in RPE1 cells expressing WT or centrinone-resistant G95L (homozygous knock-in at the endogenous locus) Plk4. (Right) Passaging assay on RPE1 Plk4 G95L cells. Data are means T SD (N = 2). (C) Lineage analysis showing the percentage of RPE1 cells with the indicated number of centrosomes arresting in each generation after centrinone addition (N = 2). Cells coexpressing centrin-GFP and H2B-RFP were initially filmed in both GFP and RFP channels to count centrosomes and monitor mother cell mitosis. Daughter cell fate was subsequently tracked by using RFP only. Arrest was the inability to enter mitosis within 48 hours of cell birth. (D) Western blot of p53 and p21 in RPE1 cells. a-tub, a-tubulin. (E) Western blot of RPE1 cells expressing control or p53 shRNA, and passaging assay of RPE1 cells expressing control or p53 shRNA after centrinone addition. Data are means T SD (N = 2).