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. 2018 Feb 5;217(2):527–540. doi: 10.1083/jcb.201704028

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© 2018 Chitale and Richly

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Figure 5. — MMSET is required for efficient NER and interacts with the NER machinery. (A) MMSET is recruited to the site of UV damage. Immunofluorescence images showing distribution of CPDs and mCherry-MMSET in U2OS cells subjected to damage through a 3-µm micropore membrane. MMSET is recruited to ≈53% of lesions 30 min after UV exposure. Bars, 5 µm. (B) DDB2 is required for UV-dependent setting of H4K20me2 by MMSET. The graph shows a quantification of the number of MMSET-LacR arrays showing H4K20me2 in the indicated knockdown cell lines cells 2 h after UV exposure (mean ± SEM). Colocalization was measured from three independent experiments, with 30 cells per experiment. Statistical significance was assessed by an unpaired t test. *, P ≤ 0.01. (C) DICER is required for UV-dependent setting of H4K20me2 by MMSET. The graph shows a quantification of the number of MMSET-LacR arrays showing H4K20me2 in cells transfected with the indicated siRNAs 2 h after UV exposure (mean ± SEM). Colocalization was measured from three independent experiments, with 30 cells per experiment. Statistical significance was assessed by an unpaired t test. **, P ≤ 0.001. (D) MMSET interacts with ZRF1 and DICER. Western blot showing HA immunoprecipitations (IP) from cells transfected with either empty plasmid or HA-MMSET.