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Supplemental Figure Legends Supplemental Figure 1. SemaB / mice have normal immune cell populations. Cells were prepared from the spleens of WT and SemaB / mice, stained with various antibodies and then analyzed by flow cytometry. Supplemental Figure 2. SemaB / T cells, B cells and DCs have normal proliferation, cytokine production and cellular interactions. A, B, Normal proliferative responses in SemaB / B cells and T cells. Splenic B cells from WT (open circles) and SemaB / (closed circles) mice were cultured with various concentrations of anti-μ or anti-cd for 36 h (A). Splenic CD + T cells from WT (open circles) and SemaB / (closed circles) mice were cultured with immobilized anti-cd3 with or without anti-cd28 for 8 h (B). [ 3 H] thymidine was added for the last 1 h. C, Th1/Th2 skewing was not impaired in SemaB / T cells. WT (open circles) or SemaB / (closed circles) naïve CD62L + CD + T cells were cultured under Th1- (left panel) or Th2- (right panel) skewing conditions for days. Then, CD + T cells were restimulated with immobilized anti-cd3 for 2 h. The cytokine concentrations in the culture supernatants were measured by ELISA. D, Normal cytokine production of SemaB / DCs. Splenic DCs (left panel) or BMDCs (right panel) from WT (open circles) or

SemaB / (closed circles) mice were stimulated with anti-cd for 8 h. The concentration of IL-12p in the culture supernatants was measured by ELISA. E, SemaB was not involved in normal MLRs between T cells and DCs. CD + T cells from BALB/c mice were cultured with irradiated allogeneic C7BL/6 WT (open circles) or SemaB / (closed circles) DCs for 6 h (upper panel). CD + T cells from C7BL/6 WT (open circles) or SemaB / (closed circles) mice were cultured with irradiated allogeneic BALB/c DCs for 6 h (lower panel). [ 3 H] thymidine was added for the last 1 h. Supplemental Figure 3. does not affect B cells, T cells and DCs in terms of proliferation, cytokine production and antibody production. B cell proliferation (A), B cell antibody production (B), T cell proliferation (C), splenic DC (left panel) and BMDC (right panel) cytokine production (D) and MLRs between CD + T cells and allogeneic DCs (E) were analyzed in the presence of (closed bars or circles) or (open bars or circles). F, did not affect DC-mediated Th1/Th2 differentiation. Naïve CD62L + CD + T cells derived from OVA-TCR Tg mice and BMDCs were cultured with OVA peptide for days in the presence of (closed circles) or (open circles). Then, CD + T cells were isolated and stimulated with

immobilized anti-cd3. The concentrations of IFN-γ (left panel) and IL- (right panel) in the culture supernatants were measured by ELISA. Supplemental Figure. affects basophils in a dose-dependent manner. BM-derived basophils were stimulated with IL-3 in the presence of various concentrations of (closed and semi-closed bars) or (open and semi-open bars) and then examined for IL- production. Supplemental Figure. SemaB / basophils have normal IL- production. WT (open bars) or SemaB / (closed bars) BM-derived basophils were cultured with IgE anti-dnp plus various concentrations of DNP-HSA for 16 h. The cytokine concentrations in the culture supernatants were measured by ELISA.

/ / 1 1 1 1 CD8 11.8 12.31 CD 2. 2.23 CD 22.93 22.13 Foxp3 1 1 1 1 B22 8.7 1 1 1.3 1.31 CD3 1 1 1. 3.7 29.26 CD11c 2.89 2.73 2.9 2.7 1 1 1 1 DX 1 1 CD21 9.3 9.2 CD23 1 1 Supplemental Figure 1

A D Splenic DC BMDC 1 9 1 6 2 2 3 onses p.m) B cell respo ( 1 3 c.p B T ce ell responses ( 1 3 c.p.m) C IFN-γ (ng/m ml) 2 16 12 8. 2 1 Anti-μ (μg/ml).1 1 1 Th1 condition onses p.m) B cell resp ( 1 3 c.p T ce ell responses ( 1 3 c.p.m) IL- (ng/m ml) 2 1.2.8...2 1 Anti-CD (μg/ml) + Anti-CD28.1 1 1 Th2 condition E IL-12p (ng/ml) T ce ell responses ( 1 3 c.p.m) nses.m) T cell respo ( 1 3 c.p. 3 2 1 1 1 Anti-CD (μg/ml) IL-12p (ng/ml) 1 1 1 DCs ( 1 3 cells/well) 1 Anti-CD (μg/ml) DCs (C7BL/6) T cells (BALB/c) DCs T cells (BALB/c) (C7BL/6).1 1 1.1 1 1 1 1 1 DCs ( 1 3 cells/well) Supplemental Figure 2

A C B cell resp ponses ( 1 3 c.p.m) responses 3 c.p.m) T cell ( 1 E T cell response es ( 1 3 c.p.m ) 6 2 2 + Anti-μ B IgM (μg g/ml) 3 2 1 3 9 2 1 D /ml) IgG1 (ng 6 3 + + + IL-12p p (ng/ml) ml) IgE (ng/ IL- IL- IL- 6 2 Splenic DC IL-12p p (ng/ml) 9 6 BMDC + 1 1 Anti-CD3 AtiCD Anti-CD (μg/ml) AtiCD Anti-CD (μg/ml) 1 1 1 DCs ( 1 3 cells/well) F IFN-g (ng/m ml) 12 8.1.1 1 IL- (pg/m ml) 3 18 12 6 2.1.1 1 Supplemental Figure 3

9 *** /ml) IL- (pg/ 6 3 N.D. + IL-3 (2 μg/ml) (1 μg/ml) (2 μg/ml) (1 μg/ml) Supplemental Figure

6 9 IL- (ng/ml) 2 IL-6 (ng/ml) 6 3 N.D. 1 1 N.D. 1 1 DNP-HSA (ng/ml) DNP-HSA (ng/ml) Supplemental Figure

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