Characteristics of immune homeostasis in <i>Nfkbiz</i>^−/− mice.

<p>(A, B) Naive CD4⁺ cells in the spleen and lymph nodes (LNs) of 8–12 week old <i>Nfkbid</i>^+/+ and <i>Nfkbiz</i>^−/− mice. (C, D) Flow cytometric analysis of IFN-γ- and IL-17-producing CD4⁺ cells isolated from the spleen (C) and LNs (D) of <i>Nfkbid</i>^+/+ and <i>Nfkbiz</i>^−/− mice at 8–12 weeks of age. Paired data were evaluated using the Student's t test. **p<0.01.</p

<i>Nfkbiz</i>^−/− mouse T cells fail to generate Th17 cells <i>in vitro</i>.

<p>(A–D) Expression of IL-17A protein or <i>Il-17a</i> mRNA (A, B) and of the Th17-related mRNAs <i>Ccr6</i> and <i>Il-17f</i> (C, D) in CD4⁺ T cells from <i>Nfkbiz</i>^+/+ and <i>Nfkbiz</i>^−/− mice, cultured for 48 h under Th0 or Th17 conditions. (E) Diagram of <i>Il-17a</i> and <i>Il-17f</i> gene conservation. Red-arrows indicate the <i>Il-17a</i> promoter, the <i>Il-17f</i> promoter, and the CNS 1, CNS 2, and CNS 3 regions. (F) ChIP analysis of H3K27Ac. Cells were cultured under Th0 or Th17 conditions for 48 h. Data shown are from one experiment that was representative at three independent experiments. (A–D) Data shown represent mean ± S.E. (n = 3). Paired data were evaluated using the Student's t test. *<i>p</i><0.05, **<i>p</i><0.01.</p

<i>Nfkbiz</i>^−/− T cells show decreased RORγt expression.

<p>RORγt expression in CD4⁺ T cells from <i>Nfkbiz</i>^+/+ and <i>Nfkbiz</i>^−/− mice, cultured for 72 h under Th0 or Th17 conditions. Data are representative of three independent experiments.</p