FIGURE 5.

Th17 cell differentiation, CD4⁺ T cell proliferation, and STAT3 phosphorylation are inhibited by hemin but not by SnPP. A, flow cytometric analysis of magnetically purified naïve T cells from spleens of BALB/c mice cultured under Th17-skewing conditions with or without hemin or SnPP (30 nmol/ml) for 3 days. Numbers in the upper right quadrants indicate the percentages of Th17 (CD4⁺IL-17⁺) cells as well as each generation of Th17 cells (undivided cells (0), generation 1 (1), generation 2 (2), and generation 3 (3)) gated on CD4⁺ T cells. B, Western blot analysis of purified CD4⁺ T cells isolated from DO11.10 mice pretreated with or without hemin and SnPP and cultured with IL-6 for 1 h. β-Actin was used as the loading control. Densitometry analysis of p-STAT3 and total STAT3 was performed by normalizing to β-actin levels (* compared with CD4⁺ T cells from untreated mice: *, p < 0.05). All results shown are representative of three independent experiments.