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. 2015 Apr 20;14(9):1413–1425. doi: 10.1080/15384101.2015.1007785

Figure 3.

Figure 3.

USP7 disruption compromises the formation of IRIF of uH2A and FK2, thereby affecting the BRCA1 and to a lesser extent the 53BP1 recruitment to DNA strand breaks. HCT116 and HCT116-USP7−/− cells were exposed to IR at 10 Gy. Nuclear accumulations of uH2A, γH2AX, FK2, BRCA1 and 53BP1 were visualized by immunofluorescence using specific antibodies. Calibration bar is 10 μm. (A) IRIF of uH2A and γH2AX. (B) IRIF of FK2 and γH2AX. (C) IRIF of BRCA1 and γH2AX. (D) IRIF of 53BP1 and γH2AX. (E) The quantitative data of IRIF of γH2AX, uH2A, FK2, BRCA1 and 53BP1 from HCT116 and HCT116-USP7−/− cells. Only the cells with 10 or more distinctive IRIF were considered IRIF positive and scored for quantitation. Mean ± SD of IRIF vs. γH2AX positive cell ratio was calculated from 4–6 microscopic fields of 3 independent experiments. (F) Magnified views of IRIF of 53BP1 in HCT116 and HCT116-USP7−/− cells exposed to IR at 10 Gy.