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. Author manuscript; available in PMC: 2015 Dec 18.
Published in final edited form as: Mol Cell. 2014 Nov 26;56(6):777–785. doi: 10.1016/j.molcel.2014.10.020

Figure 2. R-loop-induced DNA damage depends on XPF and XPG.

Figure 2

(A) P-KAP1 level in HeLa cells transfected with siXPG or siLUC 24 hours prior to transfection with siLUC or siAQR. (B) Neutral comet assay in HeLa cells treated as in Figure 2A. Scale bar represents 50 μm. (C) Quantification of comet tail moment for the experiment described in (B). a.u. arbitrary units. ****p<0.0001. (D, E) P-KAP1 level in XPG- and XPF-patient cell lines either complemented or not with the corresponding wild-type proteins, and transfected with siLUC, siAQR#2 or siAQR#3. (F, G) Quantification of percent γH2AX-positive cells in XPG- and XPF-patient cell lines either complemented or not with the wild-type or nuclease-dead proteins, and transfected with indicated siRNA. (SEM, n=3). (H) Immunostaining with S9.6 (red) and nucleolin (green) antibodies in HeLa cells transfected with siXPG or siLUC 24 hours before transfection with siLUC or siAQR. A merge of the two channels is shown, with the nucleus (stained with Hoechst) outlined. Scale bar represents 10 μm. The levels of all panels were adjusted equally in Adobe Photoshop. (I) Quantification of S9.6 immunofluorescence intensity per nucleus for the experiment described in (H), shown as box and whiskers plot. ****p<0.0001.