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. 2014 Mar 6;10(3):e1004178. doi: 10.1371/journal.pgen.1004178

Figure 6. Nucleosome acidic patch promotes RNF168-dependent DDR signaling and inhibition of DNA resection in G1.

Figure 6

(A and B) RIF1 and BRCA1 IRIF are impaired in GFP-LANA (1–32a.a.) expressing cells. Experiments were performed as in Figure 5A. Representative images are shown. CyclinA negative and high/low LANA expressing cells are indicated in the merged image. Note: CyclinA marks S/G2 cells. (C and D) Quantification of RIF1 and BRCA1/CyclinA-positive IRIF from A and B. Graphs represent values obtained from two independent experiments where foci from >100 cells were quantified. Error bars = SEM. (E) GFP-LANA (1–32a.a.) expressing cells exhibit DNA end-resection as detected by RPA accumulation at laser damage in G1 (CyclinA-negative) cells. Experiments were performed as in Figure 5E with indicated antibodies after 4 h post micro-irradiation. (F) RPA32 laser lines in CyclinA-negative cells without or with GFP-LANA (1–32a.a.) expression are indicated by yellow and green dotted lines respectively. Enlarged images from each category are shown. All cells have been laser damaged. (G) Quantification of F and G from either WT GFP-LANA (1–32a.a.) or mutant (Mut) GFP-LANA-8LRS10 expressing cells. Laser damaged CylclinA negative cells positive for LANA expression were scored for RPA laser line formation. Graph represents data obtained from >50 cells from two independent experiments. Error bars = SEM.