Figure 2. Doxorubicin (ADM)-treated apoptotic MCF-7 cells induce immature dendritic cell (iDC) maturation. Peripheral blood mononuclear cells from healthy donors were incubated with 100 ng/mL GM-CSF and 10 ng/mL IL-4 for 6 days and were differentiated into iDCs. Apoptotic MCF-7 cells were extensively washed after treatment with ADM (5 µg/mL) for 24 h and were co-cultured with iDCs for a further 24 h (iDCs+APO). Then, the CD11c-positive DC population was gated and the DC-surface expression of CD83, CD86 and HLA-DR was analyzed by FACS. As a control, iDCs were co-cultured with MCF-7 cells for 24 h (iDCs+MCF-7). As a positive control, mature DCs were generated by further stimulating iDCs with 1 µg/mL lipopolysaccharide for 24 h (iDC+LPS). GM-CSF = granulocyte/macrophage colony-stimulating factor. A, Data are reported as mean fluorescence intensity (MFI) ± SD for three independent experiments. ^*P < 0.05 vs the iDC+MCF-7 group (one-way ANOVA). B, Representative data of MFI from three independent experiments. The gray histograms represent staining with the corresponding isotype-matched control antibodies.