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. Author manuscript; available in PMC: 2014 Apr 30.
Published in final edited form as: Science. 2013 Sep 26;342(6157):447–453. doi: 10.1126/science.1237910

Fig. 4. MUC2 binds galectin-3, dectin-1 and FcγRIIB on DCs.

Fig. 4

(A) FC of CFSE-MUC2 on human DCs pre-incubated with unlabeled native MUC2, deglycosylated (dgl) MUC2, MUC2 peptide, mannan or lactose. % of MUC2 binding compared to medium alone. (B) ELISA of native or dgl MUC2 interaction with galectins. (C) CFSE-MUC2 or CFSE-dgl MUC2 binding to DCs pre-incubated with PBS, human serum albumin (HSA) or galectin-3. (D) CFSE-MUC2 binding to DCs before and after pre-incubation with a fluorescent Ab to galectin-3. (E) IFA of CD11c, galectin-3, Muc2 and DAPI in mouse PP sections. Original magnification ×5 (upper panel) and °—63 (bottom panel). (F) qRT-PCR of mRNA for galectin-3 in DC subsets from mouse PPs, SI-LP and SPL. RE, relative expression compared to Gapdh. (G) Immunoprecipitation (IP) with control or anti-galectin-3 Abs of proteins from human DCs treated without (ctr) or with MUC2 for 30 min, followed by immunoblotting of FcγRIIB, Dectin-1 and galectin-3. (H) ELISA of IL-12p70 from human DCs exposed to scrambled (ctr) or LGALS3 (galectin-3), FCGR2B (FcγRIIB) or CLEC7A (Dectin-1) small interfering RNAs (siRNAs) and cultured with or without LPS and/or MUC2 for 2 days. (I) Binding of CFSE-MUC2 to SI-LP DCs from WT, Lgals3−/−, Clec7A−/− or Fcgr2b−/− mice. Data summarize experiments with 3 donors or 3 mice from each strain (error bars, s.d.; unpaired t test, *P <0.05) or show one of 3 experiments with similar results.