Unique responses to I-IFNs by lung DC subsets determine virus replication.
A. Sorting strategy for lung DCs. Lung DCs are defined as CD45+ CD11c+ MHC-II+ Siglec-F- Ly6C- cells (gates I-IV). From gate IV individual DC subsets were further gated into CD103+ DC (gate V) and CD11bhigh DC (gate VI) for separation by cell sorting. B. qPCR analysis of Ifnar genes and signaling related genes in lung CD103+DCs (blue bars) and lung CD11bhigh DCs isolated from IFNAR +/+ naïve mice. C. qPCR analysis of ISG expression by MLN CD103+ DCs (blue bars) or MLN CD11bhigh DCs (green bars) sorted from either IFNAR -/- or IFNAR +/+ PR8-infected mice at 4 dpi. D. Expression of viral NP mRNA was determined by qPCR for individual sorted MLN DC subsets as in (C). E. CD103+ DCs and CD11bhigh DCs isolated from the lungs of IFNAR -/- or IFNAR +/+ mice during the course of PR8 infection were assayed for viral NP mRNA by qPCR. F. Total MLN cDNA obtained from PR8-infected IFNAR -/- mice, at different time points after infection, was analyzed for mRNA of the 8 segments of influenza virus (NP, HA, M, NS1, NA, PB1, PB2, PA) by qPCR. Error bars represent mean +/− SD, * p<0.05.