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. 2019 Jul 24;47(17):9160–9179. doi: 10.1093/nar/gkz639

Figure 5.

Figure 5.

CtIP recruitment to DNA damage requires interactions with the zinc finger domain of CTCF. (A) Recruitment of CtIP to mCherry-LacI-FokI-induced DSBs was evaluated by ChIP–qPCR in the CTCF-depleted FokI–U2OS cells complemented by the indicated HA fusion CTCF proteins (see Figure 1D, top). CtIP and chromatin were immunoprecipitated with an anti-CtIP antibody. qPCR was performed for the quantitative analysis of ChIP samples. Fold recruitment values were relative to those of cells without induction of DSBs. Data are means ± SD of at least three independent experiments, and all qPCR reactions were performed in triplicate; *P≤ 0.05. (B) GFP–CtIP co-localization (green) at γH2AX foci (red) in CTCF-depleted U2OS cells complemented by the indicated HA fusion CTCF proteins (see Figure 1D, top). CTCF-depleted (siCTCF) and control (siCTL) U2OS cells were subjected to laser micro-irradiation. Cells were fixed and stained with the antibodies indicated on the left. The scale bar represents 10 μm. (C) Co-immunoprecipitation of endogenous CtIP and MRE11 in CTCF-depleted (+) or control (−) 293T cells was performed with anti-CtIP antibody. Immunoblot (IB) analysis was performed with the antibodies indicated on the right.