Figure 6
From: Thymic DCs derived IL-27 regulates the final maturation of CD4+ SP thymocytes
IL27- mediated signal in CD4+ SP thymocytes.
(A) CD4+ SP thymocytes were isolated form CD11c-cre p28flox/flox and WT mice, cultured in RPMI-1640 medium with IL-27. Cells were harvested at 0, 5, 15, 30 and 60 min after stimulation, Phosphorylated and total STAT1 and STAT3 were detected by Western blotting. The cropped gels have been run under the same experimental conditions. Representative blots are shown of three independent experiments. β-actin was used as a control. (B) SP3 cells from CD11c-cre p28flox/flox mice and WT littermates were stimulated by 2 ng/ml IL-27 for 24 hours, cells were collected and the mRNA expression of IFIT1, IFIT3, IRF7 and IRF8 were evaluated (n ≥ 3). The experiments were repeated for three times and the data are shown as mean ± s.d. (C) Total RNA was extracted from FACS-purified CD4+ SP thymocyte subsets from CD11c-cre p28flox/flox mice and WT littermates. Quantitative PCR was performed to analyze mRNA expression of IFIT1, IFIT3, IRF7 and IRF8 (n ≥ 3). *p < 0.05; **p < 0.01; ***p < 0.001.
