Proangiogenic DCs are generated in vivo under conditions associated with elevated levels of extracellular adenosine. (A) Single-cell suspensions from the lungs of wild-type or ADA^−/− knockout mice were analyzed by multicolor flow cytometry. CD45⁺ cells were gated and assessed for expression of CD11b and CD11c markers. (B) Cells gated and sorted as shown in (Ai-iv) were spun on glass slides and stained with Diff-Quick to assess their morphology. Microscopy was performed with a Nikon Eclipse E600 (Nikon Instruments) using a Nikon 100×/1.4 NA oil objective. Images were captured with Olympus DP-11 digital camera system (Olympus America). Note DCs with projections in (ii) and (iv) and that cells in (iv) are bigger. Bar, 10 μm. (C) CD11b^lowCD11c⁺ cells were gated as shown in (Aii,iv) and assessed for expression of MHCII, CD86, or CD209 markers. (D) These cells were cultured for 18 hours in RPMI with 10% FBS without or with 100 μM NECA. Culture supernatants were collected, and secreted cytokines were measured by ELISA. Average values from 3 different experiments are shown. Error bars denote SE.