FIG. 4.

(A) Expression of Gal-3 in immature BM-DCs. The left panel shows Western blot analysis of Gal-3 protein expression in DCs (5 × 10⁵ cells/lane). In the right panel, gated CD11c⁺ DCs were analyzed for Gal-3 expression (bold line). The isotype control is indicated (thin line). (B) Endocytic function of Gal-3 in immature BM-DCs was assessed by flow cytometry. WT or Gal-3^−/− BM-DCs were incubated with FITC-labeled dextran, LDN-PAA, LN-PAA, or glucitol-PAA for 2 h at 4°C (thin line) or 37°C (bold line). Endocytosis is displayed as an increase in fluorescence intensity at 37°C compared with 4°C. (C) WT DCs were either left untreated or were treated with lactose (5 mM) 30 min before exposure to FITC-labeled LDN-PAA. (D) Effects of Gal-3 deficiency on DC maturation (left panel). WT or Gal-3^−/− DCs were incubated overnight with schistosome eggs, a combination of Pam₃CSK₄ plus poly(I·C), or LPS. WT DCs were left untreated or LDN-PAA or glucitol-PAA (50 μg/ml) was added 30 min before Pam₃CSK₄ plus poly(I·C) (Pam+pIC) or LPS stimulation (right panel). After an 18-h stimulation, quantification of secreted cytokines was performed by ELISA. The mean values ± SDs are presented (four experiments performed). TNF-α, tumor necrosis factor alpha.