Differential Dynamics of ATR-Mediated Checkpoint Regulators
Figure 4
UV-induced changes in mobility of checkpoint proteins. (a) U2OS cells expressing GFP-ATRIP (left panel) and GFP-Rad1 (right panel) were left untreated or treated with UV. Two hours later, soluble and chromatin-bound proteins were isolated and analyzed by western blotting using the indicated antibodies. (b–e) Hus1-GFP (b), GFP-Rad1 (c), GFP-Rad9 (d), and GFP-ATRIP (e) expressing cells were left untreated or treated with UV and analyzed by strip-FRAP (see Section 2 for technical details). (f) U2OS cells expressing GFP-Rad9 or GFP-ATRIP were left untreated or treated with UV. After 1 hour cells were analyzed by FLIP (see Section 2 for technical details). Plotted is the loss of fluorescence during continuous bleaching. (g) U2OS cells expressing GFP-Rad9 or GFP-ATRIP were locally irradiated using a 266 nm UV laser. Plotted is the association of fluorescent signal at the locally damaged area.
