Differentiation of CD1a⁻ monocyte-derived dendritic cells to CD1a⁺ cells. MoDCs were collected on day 5 of culture and sorted by flow cytometry to CD1a⁻ and CD1a⁺ populations. (A) The expression of CD1a protein in the original and the sorted populations was monitored by flow cytometry. (B) The sorted CD1a⁻ (left) and CD1a⁺ (right) cells were cultured in fresh AIMV medium supplemented with GM-CSF and IL-4 without (nonactivated) and with inflammatory cocktail (activated). The ratio of CD1a⁻ and CD1a⁺ cells in both cultures was measured on day 7 by flow cytometry. (C) Kinetics of mRNA expression during moDC differentiation measured by real-time QRT-PCR. Mean values and SD of triplicates are documented. (D) Membrane expression of CD1a protein in the moDC population monitored by flow cytometry. A typical experiment of 3 is documented.