Transfection of 293T cells and infection of human DCs was performed as described above. Cell lysates were obtained after incubation of cells with RIPA lysis buffer (Sigma Aldrich) supplemented with complete protease inhibitor (Roche) and resuspended in a total of 50 ml of Laemmli sample buffer (Bio-Rad). Crude lysates were either boiled for 10 min or incubated at 42uC for 20 min and then kept on ice. Each sample was loaded in a polyacrylamide-SDS gel, and the proteins were electrophoretically separated by conventional methods. Proteins were transferred to nitrocellulose, and blots were incubated with anti-HA, anti-FLAG, anti-Actin anti-GAPDH (Sigma Aldrich) and rabbit polyclonal antibodies anti-hSTING [41] and anti-DENV NS3 (kind gift of Dr. Andrea Gamarnik), and developed using SNAP ID detection system (Millipore), following the manufacturer's instructions. Antibody-protein complexes were detected using a Western Lighting chemiluminescence system (Perkin Elmer).