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. 2010 Oct 4;191(1):45–60. doi: 10.1083/jcb.201003034

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© 2010 Ismail et al.

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Figure 8. — EZH2 and EED are not required for BMI1 IRIF formation. U2OS cells were transfected with control shRNA (Ctr shRNA) or GFP-tagged shRNA targeted to EZH2 (EZH2 shRNA) or EED (EED shRNA). (A and B) Nuclear extracts were prepared from these cells, and EED or EZH2 levels were assayed by Western blotting. As a loading control, membranes were probed for actin. (C) 48 h after GFP-tagged shRNA transfection, cells were exposed to IR (2 Gy, 30 min). Cells were fixed and costained with BMI1 and γ-H2AX. Representative BMI1 immunostaining images are shown in C. The GFP channel identifies cells that were transfected with shRNA and coexpress a GFP marker. Bar, 5 µm.