Colocalization of (+) and (−) strand HCV RNAs with NS5A and NS3.

Huh-7.5 cells were infected with HCV at MOI = 1.5 and at the indicated times post-infection the cells were fixed and processed for strand specific RNA detection followed by immunofluorescence staining for A. NS5A or C. NS3. For 6 and 12 hpi samples, antibody signal was amplified using the tyramide signal amplification kit (TSA) as described in the materials and methods section. Scale bar is 5 μm. B. Quantitation of (A) D. Quantitation of (C). Each error bar indicates standard deviation from 25 different images. Scale bar is 5 μm. Insets represent 10 times magnification of the merged image. Solid arrows point to (+) strand RNA colocalizing with NS5A and NS3; arrowheads point to (−) strand RNA colocalizing with NS5A and NS3.