Characterization of the longer limitin protein.
(A) Recombinant limitin was treated with or without N-glycosidase F overnight. The treated proteins were subjected to Western blot probed with antilimitin under reduced conditions. (B) BC7.12 cells (1 × 104/well) were cultured in the presence of the indicated concentrations of recombinant limitin with (●) or without (○) glycosidase F treatment for 2 days. Viable cell numbers were evaluated with a hemocytometer. Similar results were obtained in 3 independent experiments. (C) Recombinant limitin was subjected to Western blot probed with antilimitin under the indicated conditions. (D) Recombinant limitin was immunoprecipitated with heparin-Sephalose in the presence of excess of chondroitin sulfate or heparin. Each precipitate was subjected to Western blot probed with antilimitin. Similar results were obtained in 3 independent experiments.
