Figure 5. SU5416 upregulates CYP1A1.

A. Spleens were harvested from mice and processed in the standard fashion. Cells were incubated for 4 hours in culture with titrating doses of SU5416 as indicated, and afterwards mRNA was measured for CYP1A1. B. SU5416 upregulates CYP1B1 and IDO. Same assay as A, but mRNA for CYP1B1 and IDO were measured. C. Upregulation of CYP1A1 mRNA is dependent on the AHR. Splenocytes from C57BL/6J or AHR^−/− mice were exposed to media, IFN-γ 100 ng/ml, or SU5416 500 nM for 4 hours. mRNA was then harvested and assayed for CYP1A1. nd represents “not detected”. *** - p≤0.001. D. IDO upregulation by SU5416 is AHR-dependent. Same assay as in C, but IDO mRNA was assessed. * - p≤0.05. * - p≤0.01. E. SU5416 induces IDO in the pDC/T cell coculture to a greater extent than TCDD. pDC/T cell coculture was utilized as described previously [25]. Culture was performed for 5 days with SU5416 500 nM, TCDD 10 nM, FICZ 100 nM, or control, at which point mRNA was harvested and measured for IDO. * - p≤0.05. *** - p≤0.001. F. SU5416 induces FoxP3 in the pDC/T cell coculture to a greater extent than TCDD. Same assay as in E, but mRNA was assayed for FoxP3. * - p≤0.05. ** - p≤0.01. G. SU5416 enhances FoxP3 expression and CD39 on naïve T cells in the presence of TGF-β. Naïve T cells were placed in culture with DMSO (1∶4×10⁴ dilution), TGF-β (2 ng/ml), or TGF-β (2 ng/ml) and SU5416 (250 nM), harvested on day 3, and analyzed by flow cytometry. All of the above figures are representative of 3 independent experiments.