Fig. 3: H2AX C-terminal linker region promotes 53BP1 IRIF.
From: Evolved histone tail regulates 53BP1 recruitment at damaged chromatin
a Representative immunofluorescence micrographs for MDC1 and 53BP1 colocalization in U2OS wildtype and H2AX KO with SFB-H2AX Y142L reconstitution, at 1 h after 10 Gy radiation. Experiment was performed independently three times with similar results. b Representative immunofluorescence micrographs of 53BP1 in U2OS RNF168 KO with GFP-H2AX overexpression (left) and H2AX KO cells with GFP-RNF8 (middle) or GFP-RNF168 (right) at 1 h after 10 Gy radiation. Experiment was performed independently three times with similar results. c Immunoblot of SFB-H2AX Y142L in U2OS H2AX KO with or without RNF8 or RNF168 ectopic expression. Experiment was performed independently three times with similar results. d Representative immunofluorescence micrographs of ionizing radiation-induced foci for γH2AX, MDC1, and 53BP1 in H2AX KO with GFP-yHTA, yHTA-L132Y, and GFP-yHTA+X-linker mutants using γH2AX, MDC1, or 53BP1 at 1 h after 10 Gy radiation. e Quantification of 53BP1 foci as represented in (d) for the indicated expression vectors. The error bars correspond to mean ± SD of three-four independent experiments. Two-tailed unpaired T test. Source data are provided as Source Data file.
