FIGURE 2.

Effects of 3BP2 suppression on differentiation of RAW264.7 cells into dendritic cells. A, shLacZ and sh3BP2 cells were cultured with GM-CSF (100 ng/ml) and IL-4 (10 ng/ml) for 6 days and then activated for 2 additional days with LPS (10 ng/ml). Dendritic-like cell morphology was examined by light microscopy. A representative field of each culture condition is shown. Scale bars, 50 μm. B, cells were cultured as described above and stained with fluorescein isothiocyanate- or phycoerythrin-conjugated antibodies against CD11c, CD80, or CD86, followed by flow cytometry analysis. The dotted line histograms represent cells stained with isotype-matched control antibodies. The data shown are representative of three independent experiments. C, cell lines were cultured in the absence of serum for 12 h before stimulation with GM-CSF (100 ng/ml) and IL-4 (10 ng/ml) for 0, 15, 30, and 60 min. The cell lysates were subjected to immunoblot analysis with antibodies against phospho-p38, -IKKα/β, -AKT, and -ERK1/2. The same membrane was stripped and reprobed with anti-ERK2.