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. 2014 Oct 23;5(10):e1489. doi: 10.1038/cddis.2014.455

Figure 4.

Figure 4

XIAP depletion enhances sensitivity of HPV+ cells to TRAIL alone or combined with bortezomib. (a) Stable XIAP and LacZ control shRNA 089 and 090 cells were generated by lentiviral infection. The efficiency of XIAP knockdown in both cell lines was assessed by western blot analysis compared with respective control cells and calculated using ImageJ software normalised to β-actin. Blots were cut and combined at the indicated line. (b) Stable 089 and 090 XIAP and LacZ short hairpin RNA (shRNA) cells were treated for 16 h with 50 ng/ml TRAIL alone (T), 2.5 ng/ml bortezomib (B) or both combined (BT). Protein levels of XIAP, caspase-3 and cleaved poly ADP ribose polymerase (PARP) were analysed by immunoblotting. (c and d) Cell viability of 089 (c) and 090 (d) XIAP and LacZ shRNA cells in the presence of 50 ng/ml TRAIL in combination with increasing bortezomib concentrations as indicated was analysed by 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay after 48 h. Bars represent mean cell viability normalised to untreated control cells and error bars indicate S.E.M. of four independent experiments. P-values were determined by two-way ANOVA comparing XIAP shRNA with corresponding LacZ control cells (**P<0.01 and *** P<0.001)