FIGURE 5. Knockdown of TNK1 shows synergy with gemcitabine.

A, BxPC3 cells were reverse transfected with siRNA against TNK1 (non-silencing and lethal siRNA controls were included) and dosed with a range of gemcitabine concentrations. Cell viability was assessed at 96 hours using Cell Titer Glo. Data was analyzed using GraphPad Prism and the average normalized viability from four replicate wells with standard error was plotted for each treatment dose. The graph is a representative of three independent experiments. B, Cell viability data was further analyzed to directly compare the viability effects of non-silencing siRNA alone, 8 nM gemcitabine alone, or non-silencing siRNA plus 8 nM gemcitabine. Student’s t-test were completed for statistical analysis, *p<0.05 relative to NS, **p<0.05 relative to NS + gem. BxPC3 cells were treated with non-silencing or TNK1 siRNA. The graph is a representative of three independent experiments. C, Silencing of TNK1 potentiates TNFα induced apoptosis of pancreatic cancer cells. BxPC3 cells were treated with non-silencing (NS) or TNK1 siRNA. Samples were incubated with 20 ng/mL TNFα for 12 hours and whole cell lysates were analyzed for cleavage of PARP by western blotting. Protein expression of β-tubulin was completed to ensure appropriate loading of all lanes. Final images were cropped to highlight relevant bands. UT: untreated, NS: non-silencing.