Figure 5

USP9X inhibition sensitizes tumor cells to various chemotherapies. (a) A549 and H1299 cells were seeded into 96 well plates (5000 cells per well) and then exposed to various combinations of inhibitors including 1 μM WP1130 and different doses of ABT-737. An XTT assay was performed 72 h after treatment. The data shown are the average of triplicate measurements for each treatment condition. Experiments were repeated three times. (b) H1299 and HCT-116 cells were seeded into 96 well plates and then exposed to various combinations of WP1130 and either SAHA or 5-FU. An XTT assay was performed 72 h after treatment. The data shown are the average of triplicate measurements for each treatment condition. Experiments were also repeated three times. (c) WP1130 enhances ABT-737 induced apoptosis. H1299 cells were treated with different concentrations of WP1130 (0, 3, 4 or 5 μM) in combination with ABT-737 (20 μM) over a 24 h period. The PARP cleavage profile in these samples was then documented via western blotting using an anti-PARP antibody. β-actin was detected as a loading control. (d) The apoptotic cell population following various WP1130 and ABT-737 treatments was stained with Annexin-V and quantified using flow cytometry. The data shown are the average of quadruplicate assessments for each condition. Experiments were repeated twice.