Fig. 5

Gemcitabine promotes the activity of NFkB, leading to an increase in uPAR promoter activation. a Luciferase activity in BxPC-3 and PANC-1 cell lines transduced with AduPARLuc, 10 or 20 MOI respectively, and treated with 5 μM of kinase inhibitor BAY 11-7082 (Calbiochem) for 1 h, and then exposed to (20 ng/ml and 250 μg/ml) of gemcitabine for 24 h. Results are expressed as light units (LU) normalized to total protein levels (RLU). b Luciferase activity of 3xκB-Luc reporter plasmid in 293T cells treated with 200 ng/ml of gemcitabine for 24 h. Results are expressed as LU normalized to β-gal activity. c Luciferase activity of puPARLuc and puPAR-NFκBmut reporter plasmid in 293T cells treated with 200 ng/ml gemcitabine for 24 h. Results are expressed as LU normalized to β-gal activity. d Luciferase activity of 3xκB-Luc reporter plasmid in 293T cells transduced with increasing doses of AduPARE1A (left panel) or AduPARTK^T (right panel) and treated or not with 200 ng/ml gemcitabine. Results are expressed as fold-change of LU normalized to total protein levels (RLU). Values represent the mean ± SEM of five independent experiments. e Luciferase activity of 3xκB-Luc reporter plasmid in 293T cells co-transfected with puPARE1A or puPARE1A-NF-κBmut, in the presence or absence of 200 ng/ml gemcitabine. Results are expressed as fold-change of LU normalized to β-gal activity (RLU). Values represent the mean ± SEM of four independent experiments