FIG 3.

MGL1 expressed by Lec1 cells retains calcium-dependent C-type lectin activity. (A) Cell surface expression of MGL1 (solid histograms) on Lec1-MGL1 (black histogram) and Lec1-ctrl cells (gray histogram) was determined by flow cytometry using an MGL-specific MAb. Unstained cells (un; dashed lines) are included for comparison. (B) Western blot analysis confirming expression of MGL in cell lysates from Lec1-MGL1 cells but not Lec-1-ctrl cells. Proteins were resolved by SDS-PAGE under nonreducing conditions, transferred to PVDF membrane, and detected using MGL-specific MAb. Data are representative of 3 independent experiments. (C) Binding of galactose-PAA to Lec1-MGL1 (black histogram) but not Lec1-ctrl cells (gray histogram) in buffer containing either 10 mM CaCl₂ (solid histograms) or 5 mM EDTA (dashed lines). Data are representative of 2 independent experiments.