DENV2 Replicon Luciferase Assay

One million K562 cells were electroporated with 3 μg of DENV2-luciferase replicon [^{38 (link)}] (provided by Jan Carette, Stanford University) in 100 μL SF Cell Line Nucleofector solution (Lonza V4XC-2012) using pulse code FF-120 (Amaxa 4D Nucleofector, Lonza). Cells were incubated for 10 minutes at room temperature following nucleofection, resuspended in 500 μL of RPMI with 7% FBS, then distributed into a 48-well plate (250 μL per well) and incubated at 37°C. At each time point, cells were lysed using the Renilla-Glo^® Luciferase Assay System (Cat #E2710, Promega) according to manufacturer suggestions, and frozen at −20°C. Samples from each timepoint were then concurrently processed using the Renilla-Glo^® Luciferase Assay System according to manufacturer instructions, and analyzed on a plate reader (Infinite M1000 Pro, Tecan).