Fig. 4: Molecular dissection of the H2AX C-terminal linker region-mediated 53BP1 IRIF formation.
From: Evolved histone tail regulates 53BP1 recruitment at damaged chromatin
a Sequence of H2AX ortholog Y to L mutants. Conserved residues are highlighted in black. Leucine substitution highlighted in purple. b Representative immunofluorescence micrographs of ionizing radiation-induced foci for 53BP1 in H2AX KO with GFP-H2AX ortholog Y to L mutant reconstitutions at 1 h after 10 Gy radiation. c Quantification of cells with >5 53BP1 foci as represented in (b) for the indicated expression vectors. The error bars correspond to mean±SD of three-four independent experiments. Two-tailed unpaired T test. d Quantification of 53BP1 foci as represented in (b) for the indicated expression vectors. Foci counts are representative of ≥25 cells from three independent experiments and line is representative of the mean. Two-tailed unpaired T test. e H2AX mutants C-terminal sequences used in (f). Residues with an alanine substitution are labeled in red. Conserved residues are highlighted in black. Leucine substitution highlighted in purple. f Representative immunofluorescence micrographs of 53BP1 in H2AX KO with GFP-H2AX 19A Y142L or GFP-H2AX 14A Y142L at 1 h after 10 Gy radiation. g Quantification of cells with >5 53BP1 foci as represented in (f). The error bars correspond to mean ± SD of three–five independent experiments. Two-tailed unpaired T test. h Quantification of 53BP1 foci as represented in (f). Foci counts are representative of ≥25 cells from three independent experiments and line is representative of the mean. Two-tailed unpaired T test. Source data are provided as Source Data file.
