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Fig. 2. Heparan sulfate proteoglycans increase in regenerating skeletal muscle fibers after barium chloride-induced damage. Transverse sections from adult TA in control conditions (A,E-F), and 4 (B), 5 (C) and 15 (D) days after barium chloride injection. Staining with anti-
-heparan sufate antibody 3G10 (A-D, red) in cross sections pre-treated with the enzyme heparitinase, suggests that heparan sulfate proteoglycans are associated with regenerating skeletal muscle fibers. As a specificity control, anti--heparan sulfate antibody staining of a section not pre-treated with heparitinase is shown (E). Hoechst 33258 staining of the same section is shown in (F). Scale bar: 50 µm. (G) On western blots with anti--heparan sulfate antibody, several bands appear to increase during skeletal muscle regeneration, and according to their electrophoretic migration pattern are probably perlecan (per), syndecan-3 (syn-3), glypican (gly) and syndecan-4 (syn-4, which appears with longer exposure times, lower inset), as is indicated at the right of the figure. Western blot detection of decorin (dcn), a proteoglycan that shows only minor variations during skeletal muscle regeneration (Casar et al., manuscript in preparation), in aliquots of the same proteoglycan-enriched samples, treated with chondroitinase ABC, is shown in the lowest inset as a loading control. (H) Electrophoretic migration pattern of the core proteins of heparan sulfate proteoglycan is similar to that observed in C2C12 myoblasts extracts with the anti--heparan sulfate antibody. Molecular mass standards are shown at the left (kDa).
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