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. Author manuscript; available in PMC: 2009 Mar 1.
Published in final edited form as: J Invest Dermatol. 2008 Oct 9;129(3):573–583. doi: 10.1038/jid.2008.276

Figure 1. Galectin-3 is expressed in bone marrow-derived dendritic cells.

Figure 1

Immature DCs were prepared from bone marrow cultures in the presence of rGM-CSF, and mature DCs were obtained by additional treatment of immature DCs with E. coli lipopolysaccharide. (a) Galectin-3 levels measured by flow cytometry. BMiDCs or LPS-matured DCs were treated with Fc-Block and directly processed with anti-galectin-3 antibody for detection of cell surface galectin-3, or fixed, permeabilized, and incubated with the antibody in the presence of Fc-Block for detection of intracellular galectin-3. Mean fluorescence intensities were obtained after corrections for nonspecific primary antibody and normalized to levels of galectin-3 relative to J774A.1, obtained concurrently at each time point. Inset, galectin-3 levels determined by immunoblotting from equivalent cell loads of BMiDCs and mature DCs as above. (b) Flow cytometry for detection of intracellular galectin-3, CD11c, and class II. (i) Galectin-3, (ii) CD11c, and (iii) class II on BMDCs summarized in (a). Unshaded histogram, antibody control; shaded histogram, galectin-3 antibody. (c) Galectin-3 detected by indirect immunofluorescence microscopy. Gal3−/− and gal3+/+ BMiDCs were activated with MCP-1 and processed for visualization of galectin-3 with polyclonal anti-galectin-3 after fixation and permeabilization. Nuclear staining with Hoechst 33342 is also shown. Images were obtained with an Olympus IX61 with × 60 objective (NA 1.2). Bar represents a distance of 10 µm.