Fig. 2— Staining of heparinase-I/III and chondroitinase-AC treated rat soleus muscle cryosections. Non-treated (a, d and g), heparinase-I/III treated (b, e and h) and chondroitinase-AC treated (c, f and i) cryosections were incubated without primary antibody (g), with anti-heparan sulphate antibody HS4C3 (a–c), anti-chondroitin sulphate antibody IO4C12 (d–f), anti-heparan sulphate stub 3G10 (h) or anti-chondroitin sulphate stub 2B6 (i). Bound antibodies were visualised using a rabbit anti-vesicular stomatitis virus antibody, followed by Alexa 488-conjugated goat anti-rabbit (a–g), or using Alexa 488-conjugated goat anti-mouse antibody (h and i; control experiment omitting the primary antibody was blank (not shown)). Anti-glycosaminoglycan antibodies stain the muscle endo- and perimysium and capillary endothelium (a and d) and this staining disappears upon enzymatic depolymerisation of the glycosaminoglycan chains in question (b and f), not when the reciprocal glycosaminoglycan is digested (c and e). Staining of heparan sulphate- and chondroitin sulphate-stubs confirms the depolymerisation of these glycosaminoglycans (h and i). Scale bar = 50 μm.