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. 2020 Nov 13;11:5775. doi: 10.1038/s41467-020-19502-5

Fig. 6. CHD7 and HDAC1 regulate DNA damage-induced chromatin dynamics.

Fig. 6

a Chromatin relaxation in U2OS cells as measured by the thickness of the photoactivated PAGFP-H2A area in cells transfected with the indicated siRNAs and expressing the indicated mCherry fusions prior to 800 nm multiphoton laser micro-irradiation. b Quantification of chromatin relaxation at 120-s post irradiation from (a). Boxplot shows the first, median and third quartiles from >40 cells from 3 to 7 independent experiments. c H4 de-acetylation at 365 nm UV-A tracks 15 min after DNA damage induction in U2OS cells transfected with indicated siRNAs and treated for 1 h with the PARP inhibitor olaparib before micro-irradiation. γH2AX is a DNA damage marker (upper panel). The mean ± SD from 16 to 31 cells per condition is shown (lower panel). d Chromatin condensation in U2OS cells as measured up to 35 min post irradiation by the thickness of the photoactivated PAGFP-H2B area in cells treated for 5 min with the HDAC inhibitor SAHA prior to 405 nm laser micro-irradiation (upper panel). Quantification of chromatin condensation is presented as the mean ± SEM of 28 cells per condition from a representative of 3 independent experiments (lower panel). e Chromatin condensation in U2OS cells transfected with the indicated siRNAs and expressing the indicated siRNA-resistant mCherry fusions as measured up to 20 min post irradiation by the thickness of the photoactivated PAGFP-H2A area prior to 800 nm multiphoton laser micro-irradiation. f Quantification of chromatin condensation at 20 min post irradiation from e. Boxplot shows the first, median and third quartiles from >40 cells from 3 to 4 independent experiments. g Chromatin condensation in U2OS cells as measured up to 35 min post irradiation by the thickness of the photoactivated PAGFP-H2B area in cells treated with the indicated siRNAs, and for 5 min with the HDAC inhibitor SAHA prior to 405 nm laser micro-irradiation. h Quantification of chromatin condensation from g is presented as the mean ± SEM of 20–25 cells per condition from a representative of 3 independent experiments. Statistical significance was calculated using the two-tailed Student’s t test (all panels). Scale bar 10 µm. Source data are provided as a Source Data file.