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. 2021 Jul 7;12:683680. doi: 10.3389/fimmu.2021.683680

Figure 4.

Figure 4

BHLHE40 deficiency increases IL-10 in naïve CD4+ T cells. (A) Timeline of generating BHLHE40-edited CD4+ T cells. (B) In vitro expansion of mock-treated and sgBHLHE40-edited naïve CD4+ T cells. Cells were collected and counted with an automated cell counter after 2, 14-day expansions with Immunocult. n = 15. (C) Representative flow cytometry histogram of CFSE dilution 3 days post αCD3/αCD28 Dynabead stimulation. Gated on live singlets (left). Cumulative CFSE proliferation data of all samples (right). (D) Cytokine gene expression. IL10 expression 6 hours after stimulation with αCD3/αCD28 Dynabeads. Expression is normalized to the house keeping gene, RPLPO. n = 6. (E) T cells were stimulated with plate-bound αCD3 and soluble αCD28, and supernatant was collected at 24h (IL-2) and 48h (IFN-γ, IL-4, IL-10) to measure cytokines by ELISA. n = 10, *p value < 0.05, **p value < 0.01, Wilcoxon matched-pairs signed rank test.