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Fig. 4 | Journal of Neuroinflammation

Fig. 4

From: Amelioration of experimental autoimmune encephalomyelitis by clozapine is not associated with defective CD4 T cell responses

Fig. 4

Clozapine promoted Treg expansion in an in vitro model of T cell differentiation. 2D2 TCRMOG35-55 splenocytes were cultured at 1 × 106 cells/well in Th differentiation medium and stimulated with MOG35-55 peptide for 72 h. Representative dot plots of (a) T-bet and IFN-γ in Th1 differentiating medium, b RORγT and IL-17A in Th17 differentiating medium, and c CD25 and Foxp3 in Treg differentiating medium gated on CD4+ T cells after 72 h of stimulation with MOG35-55 with clozapine (20 μM) or vehicle. Graphical presentation of IFN-γ+ (d), IL-17A+ (f), or CD25+ Foxp3+ (h) T cells. Graphical representation of T-bet+ (e) and RORγT+ (g) as percentage of CD4. Foxp3 expression presented as geometric MFI in all CD4 T cells after differentiating to Tregs (i). IFN-γ (j) and IL-10 (k) in the supernatant from splenocytes stimulated with MOG35-55 peptide for 72 h. ****p < 0.0001 as assessed by one-way ANOVA with Dunnett’s multiple comparison test. **p < 0.01, ****p < 0.0001 by Mann-Whitney test. Shown is a representative experiment of 4

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