The gene name Nfkbiz appears incorrectly in the figure legends. The correct gene name should be Nfkbid.The authors have provided the corrected figure legends below.
Characteristics of immune homeostasis in Nfkbid
−/− mice.
(A, B) Naive CD4+ cells in the spleen and lymph nodes (LNs) of 8–12 week old Nfkbid
+/+ and Nfkbid
−/− mice. (C, D) Flow cytometric analysis of IFN-γ- and IL-17-producing CD4+ cells isolated from the spleen (C) and LNs (D) of Nfkbid
+/+ and Nfkbid
−/− mice at 8–12 weeks of age. Paired data were evaluated using the Student’s t test. **p<0.01.
Experimental autoimmune encephalomyelitis (EAE) model in Nfkbid
−/− mice.
(A) Disease progression of EAE in Nfkbid
+/+ (n = 11–13) and Nfkbid
−/− mice (n = 9–11). (B-D) Analysis of mice 12 days after immunization. (B) Cytokine profile of CD4+ cells in draining LNs. (C, D) Measurement of IL-17A (C) and IFN-γ (D) supernatant concentrations by ELISAs (Nfkbid
+/+: n = 5; Nfkbid
−/−: n = 6), using cultured draining LNs incubated in the presence or absence of MOG peptide (10 ng/ml) for 72 h. Data shown represent mean ± S.E. Paired data were evaluated using the Student’s t test. *p<0.05, **p<0.01. (E) Histology of spinal cord specimens in EAE models. Twelve days after MOG immunization, Nfkbid
+/+ and Nfkbid
−/− mice were sacrificed and their lumber section of spinal codes were collected. Three-micrometer-thick sections were stained with hematoxylin and eosin (HE), Klüver-Barrera staining (KB) or galectin-3 (Gal-3) immunohistochemistry. Serial sections were used for HE staining, KB staining and Gal-3 immunohistochemistry. Arrowheads in HE staining and KB staining indicate the demyelinated lesions. Data are representative of 3 independent experiments.
Nfkbid
−/− mouse T cells fail to generate Th17 cells in vitro.
(A–D) Expression of IL-17A protein or Il-17a mRNA (A, B) and of the Th17-related mRNAs Ccr6 and Il-17f (C, D) in CD4+ T cells from Nfkbid
+/+ and Nfkbid
−/− mice, cultured for 48 h under Th0 or Th17 conditions. (E) Diagram of Il-17a and Il-17f gene conservation. Red-arrows indicate the Il-17a promoter, the Il-17f 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. *p<0.05, **p<0.01.
Nfkbid
−/− T cells show decreased RORγt expression.
RORγt expression in CD4+ T cells from Nfkbiz
+/+ and Nfkbid
−/− mice, cultured for 72 h under Th0 or Th17 conditions. Data are representative of three independent experiments.
Passive EAE model using adoptive T cell transfer.
Collected draining LNs from the Nfkbid
+/+ and Nfkbid
−/− mice at day 12 after MOG immunizations. LN cells were re-stimulated by MOG (10 ng/ml) after 3 days in culture, and CD4+ T cells were isolated using the CD4+CD25+ Regulatory T cell Isolation Kit (Miltenyi Biotec). Nfkbid
+/+ mice (n = 3–4/group) were intravenously injected (5 × 105 CD4+ T cells/mouse) and EAE symptoms were scored for up to 12 days. In addition, these mice received 500 ng pertussis toxin (Sigma) by i.p. injection to boost their immunological responses on Days 0 and 2. Data shown represent mean + S.E. Paired data were evaluated using the Student’s t test. *p <0.05.(TIF)Click here for additional data file.