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Zika Virus E Protein and Viral RNA Signal Quantification12-bit multichannel imageswereopened with thefreeonlinesoftware program FIJI. The cell of interest was outlined with the freehand selections tool, using the DIC channel as a guide. The threshold was set to a minimum of 900 and maximum of 4,096 for the viral staining (E protein or viral RNA). The number of individual foci was quantified with the analyze particles tool (Analyze > Analyze Particles). The size range of counted foci was 1-infinity pixels, to remove background in the quantification. Foci counts per cell were recorded for all cells in each condition (R12 cells percondition perexperiment) in a Microsoft Excel document and represented as a bar graph using the mean and SD. 59ce067264