Fig. 5. Analysis of hyaluronan and chondroitin/dermatan sulfate by FACE.
A, unlabeled glycosaminoglycans were digested with chondroitinase ABC and fragments derivatized with 2-aminoacridone, separated by gel electrophoresis, and visualized by fluorescence as described under “Experimental Procedures.” Fragments were identified by co-electrophoresis with standards. Di-HA, hyaluronan-unsaturated disaccharide; Di-0S, chondroitin/dermatan-unsulfated unsaturated disaccharide; Di-4S, chondroitin/dermatan 4-O-sulfated unsaturated disaccharide; Di-6S, chondroitin/dermatan 6-O-sulfated unsaturated disaccharide; Di-diS, chondroitin/dermatan 4,6,-O-disulfated disaccharide. B, relative abundance of chondroitin/dermatan fragments was determined by quantitative image analysis of fluorescent gels similar to those in A. The total chondroitin/dermatan fragments in each lane was normalized to 100. Error bars show the mean ± S.D. of analyses of triplicate cultures. White bars, Di-0S; black bars, Di-6S; gray bars; Di-4S. C, hyaluronan fragments were calculated as in B as a percentage of chondroitin/dermatan sulfate in the same sample.