Human Flt3L Expands Dog DCs Figure 5. DC phagocytic activity. Peripheral blood cultures were collected after 7 days in culture with recombinant canine IL-4 and GM-CSF or Ad.hFlt3L conditioned medium and incubated for additional 24 h without added cytokines in the absence (mock) or presence of CpG2216 or LPS. Activated dog DCs were mixed with 1.0 mm yellow-green fluorescent FluoSpheres for 4 hrs at 4uC or 37uC. A, Confocal microphotographs show CD11c+ and MHCII+ cells (red) containing yellow-green beads in their cytoplasm. Nuclei were stained with DAPI (blue). Scale bars: 10 mm. B, Scatter plot shows the quantification of CD11c+ DCs containing yellow-green fluorescent FluoSpheres in their cytoplasm. *p,0.05 vs mock ‘ p,0.05 vs. IL-4/ GM-CSF (Two-way ANOVA followed by Tukey’s Multiple Comparison test). doi:10.1371/journal.pone.0011074.g005 PLoS ONE | www.plosone.org 8 June 2010 | Volume 5 | Issue 6 | e11074