Fig. 5

E2F1/miR-888/APLF rewiring impairs c-NHEJ and enhances cell invasiveness. a-d PFGE assays estimating the dose-response (left) and kinetics of rejoining of DSBs (right) in (a) UMUC-3 with E2F1 knockdown (KD), (b) miR-888-5p knockdown (ZIP-888), (c) APLF overexpression, and in (d) RT-4 with APLF knockdown (KD, clones A and C) versus their controls. All PFGE results represent the mean and standard deviation calculated from six determinations in two experiments. APLF protein levels were verified by immunoblotting, using actin as loading control. Impairment of DSB repair due to APLF reduction is evidenced only in aggressive UMUC-3 BC cells with an active E2F1/miR-888-5p pathway, while APLF loss in the less-invasive RT-4 cells in the absence of miR-888-5p has an insignificant effect on DSB repair. e Invasion assays of UMUC-3 with E2F1 KD, miR-888-5p KD (left) or ectopic APLF expression versus APLF-depleted RT-4 cells (right) compared to the controls (* p < 0.05, ** p < 0.01)