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. 2014 Feb 24;11(3):232–244. doi: 10.1038/cmi.2014.3

Figure 4.

Figure 4

Cytokine defects in DKO mice both in vivo and in vitro. (a) Runx3-deficient, ThPok-deficient, DKO and WT mice were injected i.p. with 40 µg/kg of αGalCer for 2 h. Protein levels of IFN-γ, TNF-α, IL-6, IL-4 and IL-17A were then detected in the serum by ELISA. Each group of experiments was performed more than five times. *P<0.05, **P<0.01, ***P<0.001. (b) Twenty-four hours after αGalCer treatment, serum was collected and used to determine IFN-γ, IL-6 and TNF-α levels. IL-4 and IL-17A were not detected. (c) Thymic iNKT cells were sorted from the indicated mice and stimulated with plate-bound anti-CD3 (1 µg/ml) for 2 or 24 h. After stimulation, the supernatant was obtained to determine the secretion of the indicated cytokines by ELISA. The results in c are representative of four to eight independent experiments. (d) Liver MNCs were isolated and activated for 3 h by PMA and ionomycin, stained for surface markers, then intracellularly stained with Abs against IL-17A or with isotype controls. All plots are gated on αGalCer+TCRβ+ iNKT cells. Data are from three independent experiments. Ab, antibody; αGalCer, α-galactosylceramide; DKO, double knockout; iNKT, invariant natural killer T; MNC, mononuclear cell; Runx3, runt-related transcription factor 3; WT, wild-type.