Induction of CCRL2 in maturing DCs. DCs were generated in vitro from CD34⁺ bone marrow precursors. (A) Total RNA was purified from immature DC (0 hours) or DC treated for different times with 100 ng/mL LPS and analyzed by Northern blot. Ethidium bromide staining is shown. (B) Kinetics of CCRL2 and CCR7 membrane expression in DC stimulated with 100 ng/mL LPS. The graphs show the mean fluorescence intensity (MFI) of cells labeled with CCRL2 and CCR7 mAbs. (C) FACS analysis of DCs treated with 100 ng/mL LPS, 20 ng/mL TNFα, or 25 μg/mL poly (I:C) for 12 hours. Data are expressed as folds of increase of CCRL2 labeling over vehicle-treated DC. (D) CCRL2 expression in spleen CD11c⁺ DCs isolated 6 hours after intravenous injection of 25 μg/mouse of LPS. Histograms represent CCRL2 expression in WT (dotted line) and CCRL2^−/− mice (solid lines). Data are representative of 2-4 separate experiments.