Figure 1. Ezh2, Suz12, and Eed recruit to laser-induced DNA damage. U2OS cells expressing GFP-Ezh2, GFP-Suz12, or GFP-Eed were laser micro-irradiated and monitored using time-lapse microscopy. Representative images at 0, 2, and 100 s are shown after laser micro-irradiation. Accumulation of GFP-Ezh2, GFP-Suz12, and GFP-Eed on the laser damage tracks over time were quantified and plotted (n = 20). Increased fluorescence on the damage tracks is plotted over time. Ezh2 does not recruit to UV damage. (B) U2OS cells were grown on a coverslip for 24 h prior to the experiment. The coverslip was then placed below a polycarbonate membrane and exposed to UV radiation (120 J/m2). Cells were then left to recover for 20 min and then fixed with 4% PFA. Cells were immuno-stained with Ezh2 and γ-H2AX antibodies. (C) U2OS cells were transfected with GFP-Ezh2 24 h before the experiment. Cells were either treated with UV (120 J/m2), MNNG (100μM) or left untreated and fluorescent recovery after photobleaching was then performed to determine the kinetics of GFP-Ezh2. Relative intensity of the bleached region was plotted over time. (D) Ezh2 recruitment is not dependent on H2AX, ATM, or DNA PK. GFP-Ezh2 was transfected in H2AX KO, EBS, or MO59J cells 24 h before the experiment. Cells were then laser micro-irradiated and imaged over time. Representative images at three different time points are shown and quantifications (an average of 12–15 cells was plotted) of the micro-irradiated regions were shown on the right.