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Figure 3 | BMC Immunology

Figure 3

From: Increased production of pro-inflammatory cytokines and enhanced T cell responses after activation of human dendritic cells with IL-1 and CD40 ligand

Figure 3

CD40L+IL-1β-treated DC secrete IL-12 and IL-23 heterodimers A-B. Immature DC were stimulated for 24 hours with LPS+IFN-γ, CD40L+IFN-γ, CD40L+IL-1β+IFN-γ, CD40L or CD40L+IL-1β. Supernatants and control AIM-V medium were concentrated by centrifugation, then subject to immunoprecipitation with anti-IL-12p40/p70 monoclonal antibody (clone C8.6, PharMingen) and A/G agarose beads. Immunoprecipitants were separated by 10% SDS PAGE under non-reducing conditions. A. Detection of IL-12p70 using IL-12p70-specific rat mAb 20C2 (PharMingen). B. Detection using anti-IL-12 polyclonal goat Ab (R&D Systems). C. RT-PCR analysis for p19 and 18s RNA. DC were stimulated with indicated stimuli for 8 h. Total RNA was extracted, reverse-transcribed and used as template for amplification during 35 cycles. Results show ethidium bromide detection of PCR products on agarose gel (represents one of 3 separate experiments).

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BMC Immunology

ISSN: 1471-2172

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