FIGURE 3.

FACS analysis of total β-catenin protein levels in presence or absence of SB216763. To assess β-catenin levels in active Treg cells, we elected to mimic a suppression assay without radioactive isotopes. A time course was performed to trend the β-catenin intensity in Treg cells in intervals of 0, 24, 48, and 72 h. Given that GSK-3β targets β-catenin for degradation, the effect of GSK-3β inhibition on β-catenin was measured using flow cytometry with targeting FITC antibodies. Unsurprisingly, an increase in total β-catenin protein was observed in FoxP3⁺ Treg cells co-cultured with CD4⁺CD25⁻ cells and treated with SB216763. All data are representative of at least two independently performed experiments, gating on CD4⁺CD25^+hiFoxP3⁺ cells.