Research Paper Volume 4, Issue 11 pp 790—802
The helicase and ATPase activities of RECQL4 are compromised by mutations reported in three human patients
Figure 6. Recruitment to DNA double-strand breaks is not affected. (A) The recruitment of either WT or mutant RECQL4 to DNA double-strand breaks induced by microirradiation is shown for six time points (0, 10, 30, 60, 150 and 300 seconds). Damage sites are indicated by white arrows. In all cases recruitment occurs rapidly, peaks after about one minute and has largely abated after five minutes. Scale bars are 10 μm. (B) Protein recruitment quantified as the ratio of intensities at the damage site compared to the rest of the nucleus. The absolute signal from five examined cells is plotted with standard error of mean. While there is variation in absolute signal, the recruitment and retention dynamics of WT and mutants are remarkably similar. Only P466L shows a trend of accelerated release from the damage site. WT and F697L are plotted on the left y-axis, P466L and F637S on the right. (C) Immunohistochemical staining of γH2AX (green) following laser irradiation of the nucleus (DAPI, blue) to confirm the creation of double-strand breaks.