Fig. 3.

UFL1 regulates the DNA damage response. a–d U2OS cells integrated with HR or NHEJ reporter and infected with UFL1 Tet-on shRNA1 and shRNA2 virus were treated with doxycycline and subjected to the HR assay and NHEJ assay as described in the method. Data presented as mean±SD of n = 3 biological replicates. *p < 0.05, **p < 0.01. Statistical significance was calculated using Student t-test. e, f Colony-formation assay following IR was performed for U2OS cells stably expressing UFL1 Tet-on shRNA1 and shRNA2 with or without doxycycline (Dox). The data presented as mean±s.e.m of n = 3 independent experiments. *p < 0.05, **p < 0.01. Statistical significance was calculated using two-way ANOVA. g Cells expressing inducible UFL1 shRNA1 and/or reconstituted with shRNA resistant wildtype (WT) and ligase dead mutant Flag-UFL1 (DEAD) were irradiated with 2 Gy IR. Thirty minutes later, cells were lysed and blotted with indicated antibodies. h UFSP2-Flag or vector plasmids were transfected into U2OS cells. 24 h later, cells were irradiated with 2 Gy IR, and cell lysates were blotted with indicated antibodies. Source data are provided as a Source Data file