FIG 6.

RACK1 is required for the positioning of flavivirus molecular components during virus replication. Huh7 cells were transfected with NTC/RACK1 siRNA for 24 h prior to infection of DENV2 or ZIKV PRVABC59 (MOI = 1). (A) Cells were subjected to indirect immunofluorescence labeling with primary (anti-NS1 and anti-dsRNA) and secondary (mouse anti-IgG 488 and anti-IgM-555) antibodies. DAPI was used to stain nuclear DNA in the merged images. (B) The signal of both channels was thresholded to remove background noise with green channel designated as region of interest (ROI) and Pearson’s correlation coefficient (PCC) was then determined and plotted as a scatterplot (scale bar: 5 μm, inset scale bar: 2 μm, images representative of n = 3 biological replicates, colocalization was calculated for 10 NTC and 10 RACK1 siRNA treated Huh7.5 cells transfected with either DENV2 or ZIKV PRVABC59).