Figure 1.

RNF126 is recruited to the UV laser micro-irradiation-induced DNA damage stripes

A. and B. Both wild-type (WT) RNF126 and catalytically-inactive RNF126(CC229/232AA) are recruited to DNA damage stripes. U2OS cells transiently expressing GFP-RNF126 or GFP-RNF126(CC229/232AA) were irradiated with a 365-nm UV laser beam (white dashed line). Images were collected every 30 sec after irradiation and representative images are shown (A). The recruitment kinetics of GFP-RNF126 (WT) and GFP-RNF126(CC229/232AA) were assessed in terms of signal intensity at DNA damage stripes relative to the un-irradiated area in three independent experiments (B). Data represent the mean ± SD. C. Diagram depicting the domain structure of RNF126 and its truncation mutants containing the N-terminus (amino acid residues 1–100), middle region (amino acid residues 101–200), and C-terminus (amino acid residues 201–311), respectively. D. and E. U2OS cells expressing GFP-tagged RNF126 truncation mutants were subjected to UV laser micro-irradiation and the recruitment of GFP fusion proteins to the DNA damage stripes were monitored in live cells. Representative images are shown (D). The percentage of cells positive for GFP fusion protein enrichment at DNA damage stripes was determined by analyzing >100 GFP-positive cells for each GFP fusion protein (E). Data represent the mean ± SD. A two-way ANOVA was performed. ^**P < 0.01. Scale bar, 10 μm. RING, really interesting new gene; RNF, ring finger protein.