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. 2003 May;77(9):5487–5492. doi: 10.1128/JVI.77.9.5487-5492.2003

FIG. 5.

FIG. 5.

Detection of nascent HCV RNA in Huh-7 cells harboring a subgenomic replicon. 9-13 replicon cells were metabolically labeled with 5-bromouridine 5′-triphosphate in the presence of actinomycin D and then subjected to double-labeling confocal laser scanning microscopy. (a) Detection of NS5A with the polyclonal antiserum WU144 and an FITC-conjugated secondary antibody. (b) Detection of newly synthesized, bromouridine-labeled viral RNA with a MAb against bromodeoxyuridine and a Texas Red-conjugated secondary antibody. (c) The overlay demonstrates colocalization (yellow) of NS5A and nascent viral RNA. Each image covers an area of 37 by 37 μm. Negative controls included identically treated naive Huh-7 cells.