DENV enters cells via clathrin-mediated endocytosis.
(A) Fluorescent image of a LCa-eYFP-expressing cell (green) infected with DiD-labeled DENV particles (red). Scale bar is 10 µm. (B) Selected frames from a single DENV particle (red), indicated with the arrow, entering a cell via clathrin-mediated endocytosis. The numbers designate seconds post-binding. (C) Histogram of the time that DENV particles colocalized with clathrin-eYFP. (D) DENV infectivity in BS-C-1 cells in the absence or presence of 15 µM chlorpromazine. Viral infectivity was measured at 24 hours post-infection by counting the number of E protein–expressing cells using immunofluorescence microscopy. The experiment was performed in triplicate, and the bars represent the average±SD. *P<0.01. (E) DENV infection in HeLa cells expressing a dominant-negative Eps15 mutant (E95/295) or its vector control (D3Δ2). At 30 hours post-transfection, HeLa cells were infected with DENV for 21 hours and subsequently stained for expression of E-proteins. Cells were analyzed by flow cytometry, and the results are expressed as the percentage infectivity in transfected cells. The experiment was performed in triplicate, and the bars represent the average±SD. *P<0.01.