Figure 6. Simultaneous treatment with P5091 and EZH2 inhibitor induces synergistic effects in DU145. (A) DU145 cells were treated with GSK126 (5 μM), EPZ6438 (20 μM), and DZNep (0.5 μM) in the absence or presence of P5091 (2.5 μM) for 48 h. Cell lysates were immunoblotted with anti-EZH2 or anti-USP7 antibody. (B) Growth curves of DU145 cells treated with GSK126 (5 μM), EPZ6438 (10 μM), and DZNep (0.5 μM) in the absence or presence of P5091 (2.5 μM), as indicated. Viable cells were counted by trypan blue-exclusion assay every 24 h after cell seeding. (C) Wound healing assays of DU145 cells treated with GSK126 (5 μM), EPZ6438 (10 μM), and DZNep (0.5 μM) in the absence or presence of P5091 (2.5 μM), as indicated. (D) Migration assays of DU145 cells treated with GSK126 (5 μM), EPZ6438 (20 μM), and DZNep (0.5 μM) in the absence or presence of P5091 (2.5 μM), as indicated. (E) Matrigel invasion assays of DU145 cells treated with GSK126 (5 μM), EPZ6438 (20 μM), and DZNep (0.5 μM) in the absence or presence of P5091 (2.5 μM), as indicated. (F) Sphere formation assays of DU145 cells treated with GSK126 (5 μM), EPZ6438 (20 μM), and DZNep (0.5 μM) in the absence or presence of P5091 (2.5 μM), as indicated. The figure shows representative images from each cell, and the scale bar is 100 μm. Values are expressed as the mean ± SD of three independent experiments (C–F). The p value was obtained by Student’s t-test. *p < 0.05, **p < 0.01.