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. 2020 Jul 22;217(10):e20191761. doi: 10.1084/jem.20191761

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© 2020 Poholek et al.

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Figure 6. — STAT3 sustains TCR-stimulated cytokine production. (A and B) Cytokine expression in live CD4⁺ YFP⁺ cells from indicated mice analyzed by flow cytometry following stimulation with PMA and ionomycin in the presence of GolgiPlug in dLNs on day 10 and the CNS on day 16 after immunization with MOG(35–55) in CFA. (C–E) dLN cells were isolated from indicated mice on day 10 after immunization and stimulated with MOG(35–55) with or without IL-23 for 19 h, GolgiPlug was added for the final 5 h, and the percentage of YFP⁺ cells expressing the indicated cytokine was analyzed by flow cytometry. (F–H) dLN cells were isolated from indicated mice on day 7 after immunization and stimulated with MOG(35–55) with or without IL-23 for 10 h, and the indicated cytokines were analyzed by ELISA in supernatants. (I) dLN cells were isolated from indicated mice on day 7 after immunization and stimulated with MOG(35–55) with or without IL-23 for 19 h, and supernatant IL-2 was assessed by ELISA. (J and K) dLN cells were isolated from Th17^ctrl mice on day 10 after immunization with MOG(35–55) in CFA and stimulated with 10 μg/ml anti-CD3 for 14 h, GolgiPlug was added for the final 5 h, and the percentage of YFP⁺ (J) and YFP⁻ (K) cells expressing IL-17 was analyzed by flow cytometry. Data points show values for individual mice with mean ± SD, pooled from two experiments (except B and C, showing one experiment representative of three with similar results). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001, one-way ANOVA.