(A) In cells expressing the wild-type GFP-CLS, localization of Myc-tagged cyclin E SWNQ-A-S180D without (left) and with (right) fusion to the PACT domain was analyzed by immunofluorescence as in Fig. 2A. Arrowheads mark centrosomes. Inset: enlarged image of merged centrosomal area. Scale bars: 5μM.
(B) DNA synthesis requires Cdk activity at the centrosome. The cyclin E double mutant SWNQ-A-S180D deficient in both centrosomal localization and Cdk binding was assessed for restoration of BrdU incorporation in CLS-expressing cells with or without fusion to the PACT domain, as indicated. Error bars represent mean ± SD (n=5).