LLPS is required for FUS and SFPQ recruitment to DNA damage sites. (A) HeLa WT cells were transiently transfected with a FUS-GFP–expressing plasmid and incubated with either 2% 1,6-HD or 2% 2,5-HD for 30 min before laser microirradiation. Upper panel: Representative micrographs of selected time points. Lower panel: Time course of FUS recruitment. (B) HeLa WT cells were transiently transfected with a SFPQ-GFP–expressing plasmid and treated as in A. Upper panel: Representative micrographs of selected time points. Lower panel: Time course of SFPQ recruitment. (C) HeLa WT cells were transiently transfected with a FUS-GFP–expressing plasmid and incubated with 50 or 100 mM Am. Ac. for 30 min before laser microirradiation. Upper panel: Representative micrographs of selected time points. Lower panel: Time course of FUS recruitment. (D) HeLa WT cells were transiently transfected with a SFPQ-GFP–expressing plasmid and treated as in C. Upper panel: Representative micrographs of selected time points. Lower panel: Time course of SFPQ recruitment. In all graphs, data are plotted as normalized average ± SEM. Scale bars: 2 µm. Arrows indicate microirradiated area.