Quantification of WNV viral RNA in myeloid cells from WNV infected mice.

<p>Mice were treated with CLL (open bars) or PBSL (black bars) 3 days prior to footpad inoculation with WNV (1000 PFU); spleens were harvested at day 3–4 p.i., and splenocytes were enriched using magnetic beads for CD11b^-, CD11c^-, or F4/80^hi cells (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0191690#sec002" target="_blank">Methods</a>). Purified myeloid populations were obtained by cell sorting and RNA isolated for quantization via qPCR. (<b>A</b>) WNV quantities/cell indicate that in infected PBSL control mice CD11b^- DCs had significantly higher WNV RNA levels than other myeloid subsets. 1-way ANOVA (B) Population frequency determined by FACS analysis on total spleen single cell preparations. 2-way ANOVA with Bonferroni post-test (C) WNV quantity per population indicates that splenic F4/80^hi MΦs and CD11b- DCs had similar levels of WNV, while CD11b⁺ DCs and MOs had much less. 1-way ANOVA. (D) CD11b^- DCs and repopulating F4/80^hi MΦs had significantly higher WNV RNA levels per population in CLL treated mice vs. control mice. 2-way ANOVA with Bonferroni post test. Data represent the combined data from 3 independent experiments. * p < 0.05, ** p <0.01.