Figure 3
Rpp21 and Rpp29 show rapid and transient recruitment to DNA damage sites. (A) Time-lapse images show the localization of EGFP-Rpp29 at the indicated time points after laser micro-irradiation of a single region, marked by a white arrow. Graph (Right) shows fold increase in the relative fluorescence intensity of EGFP-Rpp29 at laser-microirradiated sites. Each measurement is representative of at least 10 cells. Error bars indicate SD. (B) Representative images showing IRIF enriched with endogenous Rpp29 protein. U2OS cells were exposed to IR (5 Gy), fixed after 3 min recovery and stained for DNA (blue), Rpp29 (green) and γH2AX (red). A merged image is seen on the right. Results shown are typical of three independent experiments and represent at least 15 different cells. (C) As in (A), time-lapse images showing the recruitment of EGFP-Rpp21 to laser microirradiated sites. (D,E) Graphs display the percentage of Rpp21 (D) and Rpp29 (E) deficient cells showing recruitment of EGFP-Rpp29 and EGFP-Rpp21 to damaged sites, respectively, as compared with cells transfected with control siRNA. Error bars represent the SEM from two independent experiments.
