Figure 6
From: PHRF1 promotes genome integrity by modulating non-homologous end-joining
PHD domain and SDTE motif are required for PHRF1 to DNA damage sites. (a) NBS1 was depleted in NHEJ reporter H1299 cells and then I-SceI accompanied with empty vector or HA-PHRF1 were co-transfected into H1299 cells. ChIP was conducted using anti-HA antibody at 48 h after I-SceI transfection. (b) Live cell imaging shows the dynamic recruitments of GFP-PHRF1C108A, GFP-PHRF1C186A/C189A, GFP-PHRF1S915A/T917A, and GFP-PHRF1 in the presence of TBB (CK2 inhibitor), PJ-34 (PARP1 inhibitor), and ATM inhibitors to DNA damage sites generated by laser microirradiation in U2OS cells. mCherry-NBS1 was as a control. Note that PHRF1 and NBS1 almost simultaneously appeared at DNA damage sites before 180 s after laser irradiation. Arrows indicate the laser direction through the respective nuclei. GAPDH, glyceraldehyde 3-phosphate dehydrogenase; PARP1, poly(ADP-ribose) polymerase 1
