Nef activation causes inhibition in macrophage differentiation. (A) Morphologies of TF-1-fms-Nef-ER cells cultured for 24 hours with M-CSF (100 ng/mL), M-CSF/TPA (100 ng/mL), or M-CSF/TPA/4-HT (1 μM), as indicated in each case. (B) TF-1-fms-Nef-ER cells were seeded into 6-well culture plates at a density of 2 × 105 cells/well. Cells were cultured in the presence of M-CSF, M-CSF/TPA, or M-CSF/TPA/4-HT for 12 hours or 24 hours. After culture, total cells in the wells (□) and cells that adhered to the dishes (▦) were enumerated. (C) TF-1-fms-Nef-ER cells were seeded as in panel B. Cells were cultured for 12 hours in the presence of M-CSF, TPA, and increasing concentrations of 4-HT. Cells adhering to the dishes (•) or remaining in suspension (○) were enumerated. (B-C) Error bars from triplicate assays are shown and represent SD. Results are representative of 3 independent experiments.
