Cytokine-dependence of LSECtin mRNA levels in MDDC and detection of LSECtin mRNA in ex vivo peripheral blood and thymic dendritic cells. (A) Monocytes from three independent donors were treated for the indicated periods of time with either GM-CSF, IL-4, or both cytokines, or allowed to differentiate into MDDC and further matured with lipopolysaccharide or tumor necrosis factor-α. After RNA extraction from the distinct cell types, the coding region of the LSECtin mRNA was amplified and the resulting fragments resolved by agarose gel electrophoresis. (B) THP-1 leukemic myeloid cells were treated for the indicated periods of time with IL-4 alone (left panel) or for 96 hours in the presence of IL-4 and the differentiation-inducing agent PMA (right panel). After RNA extraction from the distinct cell types, the coding region of the LSECtin mRNA was amplified and the resulting fragments resolved by agarose gel electrophoresis. (C) RNA was extracted from human myeloid peripheral blood dendritic cells from three independent donors and LSECtin mRNA detected by RT-PCR. (D) RNA was extracted from human myeloid and plasmacytoid thymic dendritic cells, thymic macrophages, or liver biopsies from three independent donors and LSECtin mRNA detected by RT-PCR. In all cases, GAPDH mRNA was amplified as a control.