Fig 3.

JEV entry and infection in neuronal cells are not inhibited by chlorpromazine treatment. (A) Neuro2a, SH-SY5Y, and Vero cells were pretreated for 30 min with chlorpromazine at the indicated concentrations and given a pulse of Alexa 488-Tf for 10 min. Cells were fixed, and transferrin uptake was quantified by flow cytometry. The averages ± SDs of measured geometric means of internalized Tf in control and chlorpromazine-treated cells are shown. (B) Neuro2a, SH-SY5Y, and Vero cells were pretreated for 30 min with 50 μM chlorpromazine and infected with JEV (MOI, 0.4) in the presence of inhibitor. Infection was scored as described in Materials and Methods. Student's t test was used to generate P values. *, P < 0.05. (C) Neuro2a cells were infected with JEV (MOI, 10) in the presence of 50 μM chlorpromazine for 1 h, and the amount of virus endocytosed was estimated by qRT-PCR of JEV positive-strand RNA. (D) Virus titers (mean ± SD) in culture supernatants at 24 hpi (MOI, 0.4) were calculated by plaque assay.