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. 2023 Apr 27;136(8):jcs260698. doi: 10.1242/jcs.260698

Fig. 6.

Fig. 6.

Effects of massive DSBs induced by Hoechst 33342 sensitization following laser irradiation. (A) DSBs were induced by irradiating Hoechst 33342-sensitized cells. Time-lapse images were acquired for γ-H2AX Fab and EGFP–Ku80 (with or without 2.5 μM NU7441) in 11-4 cells, and EGFP–ATM in AT5BIVA cells. (B) Accumulation of γ-H2AX and Ku80 in untreated cells, and Ku80 with 2.5 μM NU7441, Ku80 with 5 μM KU55933, and of EGFP–ATM (ATM) in irradiated areas in Hoechst 33342-sensitized cells (mean±s.e.m. with the total number of cells indicated as n from two replicates). (C–E) FRAP with a 488-nm laser without and with DSBs. P-values for Ku80 with NU7441 or KU55933 (vs control Ku80) at 0.5, 1, 2, and 4 s were all >0.05. (C) Time-lapse images of EGFP–Ku80 and EGFP–ATM before and after bleaching. (D) Fluorescence recovery without DSBs (P<0.0001 at 0.5 s, 1 s, and 2 s, and P=0.835 at 4 s). (E) Fluorescence recovery with DSBs (P=0.0003 at 0.5 s, P=0.002 at 1 s, P=0.005 at 2 s, and P=0.049 at 4 s). *P<0.05; **P<0.01; ***P<0.001; ns, not significant (unpaired, two-tailed Student's t-test). w/, with; w/o, without. Scale bars: 5 μm.