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. 2006 Dec 1;20(23):3238–3243. doi: 10.1101/gad.1496606

Figure 3.

Figure 3.

Recruitment of MDC1 to chromatin is required for efficient NHEJ. (A) Immunoblot detection of H2AX in HeLa cells expressing control or H2AX-specific shRNA, sh3; nonspecific band from the same gel used as loading control. (B) Effect of H2AX knockdown on MDC1 TIF formation. HeLa cells expressing control luciferase or H2AX-specific shRNA were fixed 48 h post-infection with control (β-gal) or TRF2-DN adenovirus and processed for IF (γ-H2AX [green]; MDC1 [red]; and DNA counterstained with DAPI [blue]). (C) At least 100 cells were scored for five or more telomeric MDC1 foci in three independent experiments. (D) Metaphase spreads from HeLa cells expressing control luciferase or H2AX-specific shRNA 48 h post-infection with TRF2-DN or control (β-gal) adenovirus. Telomeric signals were detected with FITC-OO-(AATCCC)3 oligonucleotide and are false-colored in green; DNA (DAPI) is false-colored in red. (E) Quantification of telomere fusions in C counted as events per chromosome end in three independent experiments. All the results were confirmed with a second independent H2AX shRNA.