The phenotype of TGF-β1–induced monocytes/macrophages and their offspring of mature DCs. Murine Lin⁻c-kit⁺ HPCs were first cultured in the presence of GM-CSF + SCF for 12 days with the addition of TGF-β1 (A). These cultured cells were then washed twice and recultured in the presence of GM-CSF + TNF for an additional 3 to 5 days (B). Cells were processed for two-color immunofluorescence analyses. Gr-1 and CD40 antigens were examined by FITC-conjugated anti–Gr-1 and CD40 MoAbs. CD86, F4/80, CD16/32, E-cadherin, and DEC-205 antigens were stained with uncoupled rat-antimouse MoAbs, followed by staining with FITC-conjugated antirat IgG. The second color was shown by a PE-conjugated Ia MoAb. Quads were set up on the isotype-matched control dot plot. Representative results from three independent experiments are shown.