Figure 2

Identification of human genes that is important in endocytosis, vesicle trafficking and signaling as well as cytoskeleton rearrangement on DENV infection using siRNA screening platform. Huh7 cells were reverse-transfected with the panel of 119 siRNAs and, after 2 days, the cells were infected with DENV at an MOI of 1. After 48 hrs post-infection, the DENV infected cells were processed for immunofluorescence staining, auto-image capturing and data analysis. The data is expressed as the percentage of antigen-positive cells and the results are shown from three independent sets of experiments. To establish a baseline of infection and transfection efficiency, cells are treansfected with a control set (top left) of siRNAs and/or transfection reagent, which include transfection lipids alone or together with "non-targeting" siRNA pool, RISC-free siRNA, siGLO (fluorescently labeled) RISC free non-specific siRNA or siRNAs targeting glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Cyclo B duplex and lamin A/C. Together with these controls permitted monitoring of transfection efficiency and cytotoxicity. Accession numbers and a brief description of the role of each gene are provided in Additional File 1.