Figure 1.

Dab2 expression in BMDC, splenic DC and human monocyte-derived DC (A and B) Dab2 expression was assessed during BM (C57 BL/6) cell-derived DC (BMDC) development. Cells developing into BMDCs were harvested on days 0, 2, 4, and 6, and Dab2 mRNA and protein levels were assessed by quantitative real-time (qRT)-PCR with Fast SYBR® Green Master Mix kit (Life Technologies) and by Western blot with rabbit anti-mouse Dab2 polyclonal antibody (Protein Tech), respectively. qRT-PCR data are shown as mean ± SD of nine samples pooled from three independent experiments. (C) Cells were harvested on days 0, 2, 4, and 6 during DC development, and assessed by flow cytometry after staining with FITC-labeled CD11c and PE-labeled intracellular Dab2 antibodies. (D) Splenic DCs were isolated from mouse (C57BL/6) spleen using CD11c⁺ isolation kit (Miltenyi Biotech) and treated with or without LPS (100 ng/mL) for 24 h. Intracellular Dab2 expression in splenic DCs was assessed by FACS (left) and Western blot assay (right). (E) Dab2 mRNA and protein expression were assessed by real-time (RT)-PCR and by Western blot with rabbit anti-mouse Dab2 polyclonal antibody (Protein Tech), respectively, during human monocyte-derived DC (hMoDC) development (left) as described in Materials and Methods . Intracellular Dab2 expression in MoDCs was also assessed by FACS (right).