Fig. 4: Analysis of H2A.X deposition in chromatin upon DNA damage induction.
a H2A.X levels in chromatin. Western blot of fractionated chromatin from WT HeLa and HCT-116 cells. Total H2A.X and core H4 histone were detected with specific antibodies. Densitometry of western blot (right panel). Total H2A.X protein levels in HCT-116 cells are represented relative to HeLa cells and normalised to H4 histone. Mean ± SD presented for n = 3 independent experiments. Paired Student’s t-test, *P-value ≤ 0.05. b Western blot analysis of H2A.X in isolated chromatin from HeLa and HCT-116 cells treated with etoposide as indicated. H4 was used as loading control. M: protein size marker. Mean ± SD presented for n = 3 independent experiments. Unpaired Student’s t-test. ns: not significant P-value >0.05, *P-value ≤ 0.05, **P-value ≤ 0.01, ***P-value ≤ 0.001, ****P-value ≤ 0.0001. c As in b but for γH2A.X. d Western blot analysis of H2A.X in isolated chromatin from HeLa and HCT-116 cells after 15 J/m2 UV irradiation. H4 was used as loading control. M: protein size marker. Mean ± SD presented for n = 3 independent experiments. Unpaired Student’s t-test ns: not significant P-value >0.05, *P-value ≤ 0.05, **P-value ≤ 0.01, ***P-value ≤ 0.001. e As in d but for γH2A.X.
