Fig. 2

Enzalutamide and USP14 inhibition synergistically suppress the proliferation of breast cancer cells. a MCF-7, MDA-MB453, MDA-MB231, MDA-MB468 and HCC1937 cell lines were plated in 96-well plates and exposed to enzalutamide (Enza; 0, 5, 10, 20, 30 μM), USP14 inhibitor (IU1: 0, 50, 75, 100 μM), or the combination of enzalutamide +USP14 inhibitor for 72 h in triplicates. Cell viability was measured after adding 20 μl MTS for 2 h. *p < 0.05, ^#p < 0.01, ^&p < 0.001 vs. each treatment alone. DM: DMSO. b The indicated breast cancer cells in cultures were treated with USP14-speicfic siRNA (si-USP14, 30 nM), enzalutamide (20 μM), or both for 72 h in triplicates and then subject to the MTS assay. *p < 0.05 vs. each treatment alone. c The indicated 5 breast cancer cell lines were treated with enzalutamide (Enza, 20 μM), IU1 (75 μM), or the combination of the two agents for 48 h before they were used for the colony formation assay in which the treated cells were re-plated in 6-well plates cultured for approximately 10 days before processed for detection of the colonies. Representative images are shown. d and (e) Cell proliferative ability was detected using Edu staining in MDA-MB453 and MCF-7 cells treated with enzalutamide (Enza, 20 μM), IU1 (75 μM), or the combination of both (IU1 + Enza) for 48 h. Nuclei were stained with DAPI. The representative images (d) and pooled data (e) from three independent experiments are shown. Scale bar, 50 μm. ^&p < 0.001 vs. each treatment alone