FRAP analysis of Cdt2 binding to sites of damage in the absence of Cdt1. FRAP analysis of Cdt2 binding to sites of damage in the absence of Cdt1. (A) Quantitative.

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FRAP analysis of Cdt2 binding to sites of damage in the absence of Cdt1. FRAP analysis of Cdt2 binding to sites of damage in the absence of Cdt1. (A) Quantitative parameters, Immobile fraction, and half-maximal recovery time of the mobile fraction (T1/2), corresponding to the FRAP curves described in Fig 1C and showing the recovery of Cdt1-eGFP, Cdt2-eGFP, PCNA-eGFP, and eGFP-NSL as a control, in undamaged and locally UV-C–damaged cells. Mean values with SD, and the number of individual cells analysed by FRAP in each case are shown. Normalization and curve fitting of individual FRAP curves for parameter estimation was performed with easyFRAP. (B) FRAP images before, during, and after photobleaching at the sites of damage. (C) Mean normalized recovery curves of Cdt1-eGFP, Cdt2-eGFP, PCNA-eGFP, and eGFP-NSL in undamaged cells. (D) Efficiency of synchronization and Cdt1 depletion of MCF7 cells. The percentage of cells expressing Cdt1 or cyclin A is shown for MCF7 cells synchronized in G1, without (siLuc) or with (siCdt1) Cdt1 depletion, as described in Fig 1D. (E) Quantitative parameters, immobile fraction, and half-maximal recovery time of the mobile fraction (T1/2), corresponding to the FRAP curves described in Fig 1D and showing Cdt2 in the presence and absence of Cdt1 in undamaged and locally UV-C–damaged cells. Mean values with SD and the number of individual cells analysed by FRAP in each case are shown. Normalization and curve fitting of individual FRAP curves was performed with easyFRAP. Akiyo Hayashi et al. LSA 2018;1:e201800238 © 2018 Hayashi et al.