Expression of ligands and E3 ligases for Notch signaling in DCs.

<p><i>A</i> and <i>B</i>, Semi-quantitative RT-PCR analyses of Notch ligands (<i>A</i>) and E3 ubiquitin ligases (<i>B</i>) in BM-derived DCs prior to LPS stimulation. Serial 2-fold dilutions of PCR products were electrophoresed in agarose gel <i>C</i>, Quantitative real-time RT-PCR analysis of ligands and E3 ligases in DCs before (<i>None</i>) and after 24 h of LPS stimulation (<i>Sti</i>). The results are representative of more than five independent experiments. <i>D</i>, Flow cytometry analysis for Notch ligands in DCs. IgG, Armenian hamster IgG isotype control; None, DCs before LPS stimulation; Sti, DCs after LPS stimulation. Results are representative of three independent experiments. <i>E</i>, Western blot analysis of Mib1 in BM culture with rGM-CSF on days 5, 7, and 9, and 24 h after LPS treatment on day9 (<i>Sti</i>). <i>F</i>, Quantitative real-time RT-PCR analysis of Mib1 in BM culture with rGM-CSF on days 5, 7, and 9. <i>G</i>, Quantitative real-time RT-PCR analysis of Mib1 in the BM precursors of classical spleen DC. MP, myeloid progenitor; MDP, macrophage and DC precursor; CDP, common DC precursor; pre-cDC, committed precursors of classical spleen DC. Results are representative of three independent experiments. Data represent mean ± SD; *, P<0.05.