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. 2010 Apr 12;38(12):e129. doi: 10.1093/nar/gkq226

Figure 1.

Figure 1.

Induction of DNA double-strand breaks by local micro-irradiation. Exponentially growing normal human diploid cells were incubated for 72 h with (A, D–K) or without (B and C) 10 µM BrdU. After UVC irradiation, cells were incubated for 30 min, fixed with 4% formaldehyde, permeabilized with 0.5% Triton X-100, and incubated with anti-phosphorylated histone H2AX at serine 139 as described in ‘Materials and Methods’ section. The primary antibody was detected by Alexa488-conjugated anti-mouse IgG antibody. (A) Unirradiated cells. (B–D); Cells were exposed to 30 J/m2 of UVC with (C) or without (B and D) 5-µm pore membrane. (E–H); Cells were exposed to various doses of UVC with 5-µm pore membrane. (E) 5 J/m2, (F) 10 J/m2, (G) 20 J/m2, (H) 30 J/m2. (I-K); Cells were exposed to 30 J/m2 of UVC through membranes with different pore sizes. (I) 2-µm pore membrane. (J) 3-µm pore membrane. (K) 5-µm pore membrane. Bar indicates 2 µm.