Figure 6.

Effect of 66E2 on dengue virus infection. (A) Immunofluorescence microscopy showing DENV infection of Huh7 cells. Huh7 cells were infected with DENV-2 at 1MOI. At 48 hpi cells were fixed and stained with anti-dengue virus antibody (ab9202) and Alexa 488 anti-mouse antibody (green). Nuclei were stained with DAPI (blue). About 40% of the cells are infected. (B) Huh7 cells were infected with DENV-2 virus (at 1 MOI) in the presence of indicated concentrations of 66E2 and CMC. At 48 hpi total RNA was extracted and reverse-transcription quantitative PCR analysis was performed to determine the levels of DENV replication. Results are expressed as the percentage relative to DMSO treated control. (C) ELISA to detect levels of DENV core protein produced inside Huh7 cells infected with DENV-2 virus in the presence of 66E2 and CMC. Uninfected Huh7 cells were used as control. Results are expressed as relative percentages to DMSO treated control. For both (B and C), results are representative of three independent assays performed in triplicates. The % mean is shown above the bars and the error bars are standard deviations. (D) Huh7 cells were infected with DENV-2 at 1 MOI. At 48 hpi, total RNA was extracted from the media and viral RNA copy number/ml of the media in the presence of 10 μM of 66E2 and CMC was estimated and plotted against the compounds used. DMSO treated cells were used as control.