Fig. 6.

Anti-hensin antibody Western blots of OptiPrep density gradient fractions of LD conditioned medium incubated with galectin-3 in the presence or absence of lactose and GCT. A: 1 ml of concentrated conditioned medium from LD cultures (100 μg/ml) was placed on 11 ml of 5–40% OptiPrep density gradient medium and ultracentrifuged, and TCA-precipitated proteins from each 1-ml fraction were examined by Western blotting with anti-hensin antibody. Similar density gradient analyses of standard proteins γ-globulin (158 kDa), catalase (232 kDa), and thyroglobulin (670 kDa) were performed, and their locations in the different fractions of the density gradient are marked at bottom. The medium shows no hensin oligomerization. B: an anti-hensin Western blot of density gradient fractions of conditioned medium incubated with 20 μg/ml (0.7 μM) galectin-3 with oligomerization to fraction 12. C: anti-hensin Western blot of density gradient fractions of conditioned medium samples incubated with both galectin-3 and lactose; the lactose prevented hensin oligomerization. D: anti-hensin Western blot of the density gradient fractions of conditioned medium incubated with 0.7 μM (10 μg/ml) recombinant GCT alone; there was no hensin oligomerization.