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Fig. 3. Histone H1 binds specifically to perlecan secreted by myotubes. A total
proteoglycan-enriched fraction obtained from the conditioned media of myotubes
was incubated with histone H1 immobilized on nitrocellulose membranes and then
eluted with 8 M urea. Total, unbound and histone-H1-bound fractions were
incubated with heparitinase and separated by SDS-PAGE. Western blots were
performed using (A) an anti-
-heparan sulfate monoclonal antibody to
reveal the core proteins of heparan sulfate proteoglycans and (B) specific
anti-mouse perlecan antisera, and detected by ECL. The migration of perlecan
(per) and glypican (gly) are indicated on the right, and molecular weight
markers on the left of each panel.