Expression of MADDAM mRNA during the differentiation process of CD34⁺ cells into DCs in vitro.

CD34⁺ cells were cultured up to 15 days with FCS, stem cell factor, TNF-α, and GM-CSF (A). Total RNA was isolated and Northern blot analysis was performed as described in “Materials and methods.” As a control for RNA loading, the membrane was rehybridized with an 18S rRNA-oligonucleotide and MADDAM signals were normalized to the corresponding 18S signals. In parallel, flow cytometry analysis was performed with anti-CD1a-PE, anti-CD14-FITC, and isotype control antibodies at day 8 and day 15 of culture. The median fluorescence and the number of positive cells are shown (B).