Detection of ANDV infection of primary human dendritic cells. (A) Vero E6 epithelial cells and human immature DCs infected with ADNV (strain CHI-7913). ANDV-nucleocapsid (N) protein detected by IFA by incubating ANDV-infected with a mouse anti-ANDV N MAb revealed by an FITC-conjugated anti-mouse IgG Ab (Green, right column) while the corresponding cell nuclei were stained with 4',6-diamidino-2-phenylindole (DAPI, blue, left column). Negative control (mock) cells were incubated with supernatant from uninfected Vero-E6 cells. (B) Total RNA was extracted from uninfected LPS-Pulsed DCs (lane 1), ANDV-infected LPS-pulsed DCs (lane 2), uninfected iDCs (lane 3), ANDV-infected iDCs (lane 4), ANDV-infected Vero-E6 cells (lane 5), and used as template in a RT-PCR reaction designed to specifically amplify the viral S RNA. This assay also included a negative RT-PCR control (lane 6). MW is a molecular weight marker (1 Kb, Fermentas, Burlington, Canada). (C) DCs generated from primary monocytes, recovered from four healthy donors, were incubated with ANDV (lanes 1, 4, 7, 10), UV-irradiated ANDV (lanes 2, 5, 8, 11) or pulsed with LPS (lanes 3, 6, 9, 12). Total RNA was extracted from cell supernatants and used as a template in a RT-PCR reaction designed to specifically amplify the viral S RNA. MW is a molecular weight marker (100 pb, Fermentas).