Fatty acids treatment

Fatty acids (FAs) were purchased from Cayman and diluted in ethanol for all FAs [Palmitic acid (PA, C16:0), stearic acid(C18:0), palmitoleic acid (C16:1), oleic acid (OA, C18:1n-9), linoleic acid (LA, C18:2n6c), cis-8,11,14-eicosatrienoic acid (C20:3n6), cis-11,14,17- eicosatrienoic acid (C20:3n3), erucic acid (C22:1n9), eicosapentaenoic acid (EPA, C20:5n3), nervonic acid (C24:1n9), docosahexenoic acid (DHA, C22:6n3) and arachidonic acid (AA, C20:4n-6)] . The FAs-free medium was contained RPMI-1640 basic medium, 1×ITS medium supplement (Sigma) and 20 ng/ml GM-CSF without FAs supplement. Before addition to the culture medium, all FAs were conjugated to bovine serum albumin (BSA, fatty acid-free, Sigma) at the indicated concentration with FAs-free medium containing 1% BSA. The fatty acids were incubated with 1% BSA for 2 h before adding to DC. The concentration of saturated, monounsaturated, and polyunsaturated fatty acids in ITS medium was 20 µM. Half of the medium was discarded and replaced with fresh DC medium every other day. On day 6, DC were infected with the virus for 24 h and then analyzed by FACS.