B16F1-HyBNAR mouse melanoma cells are sensitive to human IFN-Is.

<p>(A & B) Mouse B16F1 melanoma cells were stably transfected with either the HyBNAR double transgenic construct, or a control GFP vector and tested for their anti-proliferative dose response to Hu-IFNα2, Hu-IFN-YNS and for comparison, mouse IFNβ. (A) HyBNAR transfected B16F1 cells were responsive to both human and mouse IFN-Is (EC50 values shown in parentheses) (B) In the GFP-transfected B16F1 control cells however, the human ligands only promoted a minor loss in proliferation and when administered at very high concentrations. (C) B16F1-HyBNAR and non-transfected control cells were incubated with an increasing dose of both human and mouse IFNβ. After 45 minutes incubation the cells were lysed and analyzed by Western Blot detection for phosphorylated STAT1. Total STAT1 measurements are shown as control. (D). B16F1-HyBNAR cells were treated for the indicated time-points with 100 pM human and mouse IFN-Is. Measurements of the IFN-response gene MX2 were performed by qPCR analysis. Relative fold-change was determined in comparison to untreated cells and normalized using the reference gene HPRT1. Error bars represent standard error of duplicate measurements.