Figure 2.
Cell-cycle dependent foci formation by UV micro-irradiation. Exponentially growing normal human diploid cells were incubated for 72 h with (B) or without (A) 10 µM BrdU. Cells were washed with PBS and exposed to 30 J/m2 of UVC through 5-µm pore size membrane. Then, they were incubated for 30 min, fixed with 4% formaldehyde, permeabilized with 0.5% Triton X-100, and incubated with rabbit anti-phosphorylated histone H2AX at serine 139 and mouse anti-RPA antibodies, which were detected by Alexa594-conjugated anti-rabbit IgG antibody and Alexa488-conjugated anti-mouse IgG antibody, respectively. Bar indicates 2 µm.