Figure 4.

The Endo180ΔEx2–6 mutant protein retains C-type lectin activity. Membrane preparations from +/+ and ΔEx2–6/ΔEx2–6 mouse embryonic fibroblasts were loaded onto 2-ml N-acetylglucosamine–sepharose columns. The columns were washed six times with 1 ml of loading buffer containing 25 mM Ca²⁺ and were eluted seven times with 1 ml of elution buffer containing 10 mM EDTA. Fractions were trichloroacetic-acid-precipitated, resolved by SDS–polyacrylamide gel electrophoresis using 10% gels and analysed by western blotting using the anti-Endo180 antibody followed by horseradish-peroxidase-conjugated anti-rabbit Ig.