Wnt3a and Wnt5a upregulate IDO expression and activity in DCs. A. Wnt5a induces durable IDO expression by BMDCs based on Western blot analysis at 24 and 48 hrs. Representative of 3 independent experiments. UT, untreated. XAV939, β-catenin inhibitor. B. Both Wnt3a and Wnt5a activate the IDO promoter in COS7 cells transiently transfected with a IDOprom-luciferase reporter plasmid. Performed in triplicate. Representative of 2 independent experiments. P value based on a one-way ANOVA. C. Wnt5a induces DC-derived IDO enzymatic activity based on kynurenine detection by HPLC. Cumulative data from 3 separate experiments. P value based on one-way ANOVA. D. Wnt3a and Wnt5a Stimulate β-catenin Activity in Primary TCF/Lef1-EGFP DCs in vitro. Representative of 2 independent experiments. E. Wnt3a and Wnt5a Induce β-catenin-IDO promoter binding in Primary DCs. Performed after 24 hrs of stimulation. IFN-γ induction of STAT1 binding to the IDO promoter serves as positive control. Bottom, Western blot generated from same samples. *P < 0.05. P value based on one-way ANOVA. F. β-catenin immunofluorescence microscopy confirms differential kinetics of Wnt3a- and Wnt5a-induced signaling in DCs. Bottom, examples of β-catenin nuclear translation in response to Wnt3a and Wnt5a (100x). 10 fields counted in triplicate. Representative of 2 independent experiments. All data is mean ± SEM.