Silencing Nucleolin Expression for Influenza Infection

A pool of ON-TARGETplus SMARTpool siRNA duplexes specific for human nucleolin (si C23), siRNA J-003854-05: 5′-GCAAAGAAGGUGGUCGUU-3′, siRNA J-003854-06: 5′-GAUAGUUACUGACCGGGAA-3′, siRNA J-003854-07: 5′-CAAAUCUGCUCCUGAAUUA-3′, siRNA J-003854-08: 5′-GAAAGAAGAUGAAGUUUGA-3′ (from Dharmacon, ThermoFisher Scientific) were used to silence endogenous nucleolin expression, as previously shown83 (link). Transfection was performed using Oligofectamine (Invitrogen), as previously described60 (link). Two rounds of siRNA transfection were performed – with a 24 h interval - to achieve a sufficient level of nucleolin silencing. Twelve hours after the second transfection, cells were subjected to H3N2 infection at the indicated MOIs and incubated for different time periods.