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. 2013 Dec 9;203(5):785–799. doi: 10.1083/jcb.201305109

Figure 5.

Figure 5.

CP110 can suppress centriole overduplication driven by the overexpression of centriole duplication proteins. (A) Quantification of centriole overduplication (percentage of prometaphase cells with >2 Asl-positive centrosomes) in WT and CP110Δ third instar larval brains overexpressing GFP-DSas-6 or GFP-Sak; n (total cells; total brains): 482;15, 742;14, 575;9, 695;12, 472;10, and 695;13, respectively (order as shown in graph). (B) Quantification of centriole overduplication in WT, CP110L-GFP–, or CP110S-GFP–overexpressing third instar larval brain cells with or without GFP-Sak overexpression; n (total cells; total brains): 509;9, 333;6, 374;7, 772;9, 605;11, 527;9, 1440;28, and 1888;29, respectively (order as shown in graph). (C) Quantification of centriole overduplication (percentage of cells with >4 Asl-positive centrioles) in WT and CP110Δ primary spermatocytes overexpressing the centriole duplication proteins Asl-GFP, GFP-DSas-6, GFP-Sak, GFP-Ana2, and DSas-4-GFP (as indicated); n (total cells; total testes): 317;16, 325;16, 534;22, 244;17, 250;6, 500;7, 304;5, 318;11, 506;10, 626;15, 456;11, and 161;5, respectively (order as shown in graph). Note that the apparent overduplication of centrioles in CP110Δ cells is almost certainly due to centriole mis-segregation caused by the premature separation of the centrioles, as we observed an almost equal number of cells with too few centrioles. (D) Quantification of centriole overduplication (percentage of centriole duplication events in which one centrosome divided to form more than two centrosomes) in WT or CP110Δ syncytial embryos expressing either Asl-GFP+RFP-tubulin, GFP-DSas-6, GFP-Ana2, or DSas-4-GFP; n (total centriole duplication events; total centriole duplication cycles): 669;36, 1105;49, 1298;43, 509;23, 311;7, 361;12, 301;5, and 422;13, respectively (order as shown in graph). (E and F) Panels show stills from movies of WT (E) and CP110Δ (F) syncytial embryos expressing Asl-GFP and RFP-tubulin. Time (min:sec) is shown in top right corners. Note how a very small Asl-GFP dot that can organize MTs (arrow) is present in the CP110Δ embryo close to the two centrioles that have just divided (arrowhead; t = 0:00). This dot increases in brightness over time, but does not initially divide when the other centrioles in the embryo divide (t = 14:39). This dot does divide, however, during the next round of division (t = 34:35). Note that an average percentage of centriole overduplication was calculated for each brain (A and B), each testes (C), or embryo (D), and the number of brains, testes, or centriole duplication cycles was then used as the sample number for statistical analysis. Bar: (E and F) 5 µm.