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. 2013 Sep 5;9(9):e1003585. doi: 10.1371/journal.ppat.1003585

Figure 3. Microtubules are required for RNA delivery into the cytoplasm and viral infectivity but not membrane fusion.

Figure 3

Cells were pretreated with 20 µM nocodazole (a microtubule polymerization inhibitor) for 1 h before treatment with R18-labeled JE-VLPs or YFV. (A) DIC micrograph of the treated cells showing that nocodazole has no toxic effect on Vero cells although the cell morphology is altered. (B) JE-VLPs (200 µl at 17 pM) and (C) YFV (MOI = 0.1–1) fused normally in Vero cells in the presence of 20 µM nocodazole, as indicated by the R18 fluorescence dequenching profiles of three representative tracked particles. (D) qRT-PCR nucleocapsid delivery assay showing that nocodazole reduced YFV nucleocapsid delivery into the cytoplasm of Vero cells by approximately 80% relative to untreated cells. Error bars represent the standard error of the mean (SEM) of three experiments. (E) Plaque assay showing that nocodazole treatment inhibited YFV replication, reducing the number of viral plaques by >97%. See also Movie S2.