Fig. 3

Lack of Gαq impairs ability of Bregs to suppress CD4⁺ T-cell proliferation. CFSE-labeled WT CD4⁺CD25⁻ T cells cocultured (1:1) with sorting-purified WT Bregs, Gnaq^−/− Bregs, or WT Tregs in presence of Mouse T-Activator CD3/CD28 Dynabeads™ for 72 h. Percentage of proliferated CD4⁺ T cells detected by flow cytometry (a, b), and total number of T cells also investigated (c). d, e Culture cells also harvested to analyze viability of T cells. Cells labeled with anti-mouse CD4, Annexin-V, and 7-AAD, and then analyzed by flow cytometry. Results represent mean ± SD per group (n = 5 mice/group). Data representative of three independent experiments. Student’s t test analyzed statistical difference. *p < 0.05, **p < 0.01, ***p < 0.001. Breg regulatory B cell, CON control, Treg regulatory T cell