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. 2022 Dec 17;30(2):407–416. doi: 10.1038/s41418-022-01092-y

Fig. 4. ChAT is a key determinant of Th17 cell pathogenicity.

Fig. 4

A Representative flow cytometric analysis of ChAT-GFP expression by CD4+ splenocytes polarized towards the Th17 cell fate through stimulation with plate-bound anti-CD3 plus anti-CD28 in the presence of either HM (TGFβ + IL-6), HM + IL-23, PM, or PM + TGFβ. Data are representative of at least two independent experiments. B. Quantitation of the data shown in A. Each data point represents one biological replicate (mean of three technical replicates). C EAE clinical scores of Rag1-deficient recipients transplanted with in vitro-generated and sorted 2D2 ChAT-GFP+ (green, n = 8) or 2D2 ChAT-GFP- (black, n = 7) Th17 cells. Disease severity was scored daily. Data are the mean score ± s.e.m. and are representative of three experiments. D Percent survival of the mice in C. E qPCR determination of mRNA levels (relative to Actin) of the indicated genes in ChAT-GFP+ and ChAT-GFP- splenic T cells polarized towards the Th17 fate by incubation in PM for 6 days. Data are the mean ± s.e.m. of 5 biological replicates and representative of 2 experiments.