Figure 6
RNase A treatment of cells disrupts the recruitment of EGFP-Rpp29 and EGFP-Rpp21 to laser microirradiated sites. (A,B) Representative time-lapse images (left) of mock- and RNase A-treated U2OS cells showing the localization of EGFP-Rpp29 (A) and EGFP-Rpp21 (B) at the indicated times after laser-microirradiation of a single region marked by white arrow. Results shown are typical of three independent experiments and represent at least 30 different cells. Graphs (middle) show the increase in the relative fluorescence intensity of EGFP-Rpp29 and EGFP-Rpp21 at laser-microirradiated sites in mock and RNase A-treated cells. Each measurement is representative of at least 10 cells. Error bars indicate SD. Graphs (right) display the percentage of RNase A-treated cells showing subcellular distributions of EGFP-Rpp29 and EGFP-Rpp21, as compared with those seen in untreated cells. Error bars depicts the SEM from two independent experiments. P-values were calculated by two-sided Student’s t-test relative to DMSO; **p < 0.01. (C) As in A, except for that the laser microirradiation was applied on U2OS cells expressing MonomerRed-PARP1 (MR-PARP1).
