Fig. 4

Silencing of KDM5B markedly reduces the migration and invasive potential of TNBC cells. a, b KDM5B expression in MDA-MB-231 mammosphere, cells stably expressing KDM5B vector or shKDM5B assessed by western blot analysis and RT-PCR. GAPDH serves as loading controls. c, d MDA-MB-231 control vector or KDM5B-depleted cells were subjected to a wound healing in vitro migration assay as described under Materials and methods. Representative photomicrographs at indicated time points from three independent experiments, each performed in triplicate wells, are shown. Magnification: × 20. The cells were allowed to migrate after wounding for 12 h. The extent of wound recovery was determined by measuring the distance between migrating cellular fronts at 5 randomly selected points and finding the average. Migration was significantly inhibited in shKDM5B-expressing MDA-MB-231 cells as compared with that in control wild-type cells. Column: Mean of three experiments; bar: standard error, p-value was determined by student’s t-test (*P < 0.05; ***P < 0.001). e, f KDM5B ablation in MDA-MB-231 cells significantly attenuated invasion of shKDM5B MDA-MB-231 cells as compared to control wild-type and vector cells. Column: Mean of three experiments. Bar: standard error P-value was determined by Student’s t-test (***P < 0.001). KDM5B, lysine specific demethylase 5B protein; shRNA, short hairpin RNA