Figure 1From: Melanoma-derived Wnt5a conditions dendritic cells to promote regulatory T cell differentiation via the upregulation of indoleamine 2,3-dioxygenase: novel pharmacological strategies for augmenting immunotherapy efficacyWnt3a 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.Back to article page