Figure 6: Impaired proliferation and cytokine production of Orai1/Orai2-deficient T cells in vitro.

(a) CD4⁺ T cells isolated from WT, Orai1^fl/flCd4cre, Orai2^−/− and Orai1^fl/flOrai2^−/−Cd4cre (DKO) mice were stimulated with plate-bound anti-CD3/CD28 for 4 days in vitro with or without 50 U ml⁻¹ recombinant human IL-2 (rhIL-2). Proliferation was analysed by CFSE dilution using flow cytometry; mean fluorescence intensity (MFI)±s.e.m. of CFSE from two to three experiments is shown within the histograms. Additional time points are presented in Supplementary Fig. 6. (b,c) Impaired T_H1, T_H17 and T_H2 cytokine production in Orai1- and Orai1/Orai2-deficient T cells. (b) T cells isolated from WT, Orai1^fl/fl Cd4cre, Orai2^−/− and DKO mice were differentiated for 3 days under T_H1-, T_H17- and T_H2-polarizing conditions and intracellular IFNγ, IL-17 and/or IL-4 was analysed by flow cytometry after restimulation with phorbol myristate acetate (PMA)/ionomycin for 6 h. (c) Quantification of IFNγ, IL-17- and IL-4-positive T cells as shown in b; mean±s.e.m. from three independent experiments (T_H1 and T_H17) or two experiments (T_H2). *P<0.05; **P<0.005; ***P<0.001 in (c) using unpaired Student’s t-tests.