Fig. 6

Esketamine inhibits Th17 differentiation via suppression of STAT3 activation. (A) Western blot analysis of p-STAT3 and total STAT3 levels in CD4 + T cells from the spleen of TBI mice treated with DMSO or esketamine at 40 µg/ml and 80 µg/ml. Quantification of the p-STAT3/STAT3 ratio is shown on the right (n = 3, F = 229.5). (B) Western blot analysis of p-STAT3 and STAT3 in Th17 cells differentiated from naive T cells. Quantification of the p-STAT3/STAT3 ratio is shown on the right (n = 3, F = 133.7). (C) Immunofluorescence images showing STAT3 localization in Th17 cells treated with DMSO, 40 µg/ml, or 80 µg/ml esketamine. Quantification of nuclear STAT3 intensity is shown below. Scale bar = 50 μm (F = 231.8). (D) Relative mRNA expression levels of Th17-associated genes (IL17A, IL17F, and IL23R) in CD4 + T cells treated with DMSO, esketamine, or esketamine combined with colivelin (a STAT3 activator) (F = 27.22, 33.93, 26.91 respectively). Statistical significance: Data are represented as mean ± SD of three independent experiments, *p < 0.05, **p < 0.01, ***p < 0.001