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. 2023 Jan 19;14:1058267. doi: 10.3389/fimmu.2023.1058267

Figure 1.

Figure 1

Eomes induces IL-10 production in CD4+ T cells in vitro. In vitro stimulation of splenocytes with soluble Anti-CD3 with or without IL-12 for 48 hours, cells were analyzed by RT-qPCR (B) or flow cytometry (C-F). (A) Table of Tbx21 alleles corresponding to genotypes with + = Tbx21 (encoding for T-bet), E = Eomes, - = null allele. (B) RT-qPCR of Tbx21 and Eomes in relation to house-keeping genes GAPDH and HPRT after stimulation with Anti-CD3 and IL-12. Indicated genotypes were further normalized to Tbx21+/+ (wild type), set as 100. (C) Intranuclear staining of T-bet and Eomes after stimulation with anti-CD3 and IL-12. Representative Dot plots are shown. (D) Mean fluorescence intensities (MFI) of T-bet, Eomes and mCherry. (E) Surface staining for CD44 and intracellular staining (F) for TNF, IFN-γ and IL-10 after 4 hours of stimulation with PMA, Ionomycin and Brefeldin (A) Statistical analysis: *=p < 0.05; **=p < 0.01; ***=p < 0.001; ns, not significant; Error bars denote mean + SEM. n=3.