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. Author manuscript; available in PMC: 2008 Oct 5.
Published in final edited form as: Cell. 2007 Oct 5;131(1):33–45. doi: 10.1016/j.cell.2007.08.017

Figure 4. T-bet regulates production of TNF-α in DCs.

Figure 4

T-bet is expressed in colonic DCs. (1) CD11c staining RAG2-/- colonic mucosa, 200x. (2) Serial section of (1) stained with anti-T-bet Ab, 4B10, 200x. (3) Sorted colonic, mouse DC stained with anti-T-bet Ab (red) and DAPI (blue), 1000x. (4) Human colonic DC, S100 (pink), anti-T-bet (brown), 1000x. (B) Loss of T-bet expression in BMDCs results in increased production of TNF-α.

T-bet binds the TNF-α promoter. (C) qPCR of mouse bone BMDCs ChIP samples. Primer sets A and B, approximately 500 and 1200 bps upstream of the TSS. A schema of the promoter showing T-box consensus sites and the location of primers used. Data are the mean of 3 exp (D) qPCR of human myeloid DC ChIP samples, one representative exp of 3. (E) T-bet negatively regulates TNF-α gene transcription. RAW cells transiently cotransfected with a 3kb TNF-α promoter luciferase construct, a constitutively active Iκκβ mutant construct to activate TNF-α transcription, a Renilla construct for normalization of transfection efficiency, and varying concentrations of a full length T-bet cDNA construct or control plasmid. TNF-α promoter activity is shown as a function of relative luciferase activity. Data are representative of 3 independent exp.