Fig. 3.

ROCK2 but not ROCK1 inhibition leads to down-regulation of STAT3 phosphorylation, IRF4, and RORγt expression. CD4⁺ T cells were treated with indicated doses of KD025 (A and B) or 10 μM of KD025 (C and D), and then stimulated by anti-CD3/28 mAbs, IL-1β, and TGF-β for 48 h (A, C, and D) or for 1 h (B). Nuclear (A) and cytoplasmic (B) extracts were prepared and analyzed by Western blot. ChIP assays were performed with normal rabbit IgG, anti-RNA pII, STAT3, and histone H3K27 acetylation (H3K27Ac) antibodies (C and D). Binding of these transcription factors to the sites was determined by quantitative PCR and plotted as fold enrichment compared with IgG. One representative of three different experiments is shown.