FIGURE 5.

Effect of EXTL2 overexpression on HS chain length. A and B, ³H-labeled (A) or ³⁵S-labeled (B) glycosaminoglycans were purified from the cell surface/extracellular matrix of HEK293 cells stably transfected with EXTL2, as described under “Experimental Procedures.” The resulting labeled polysaccharides were digested with chondroitinase ABC and subjected to chromatography on a Superose 6 column. The retarded component resulting at 20–23 ml corresponds to material that is degraded by chondroitinase ABC treatment. Similar results were obtained from four independent transfections with different overexpression levels. The elution positions of molecular mass standards derived from heparin (8.6 kDa) and hyaluronan (19, 30, and 43 kDa) are indicated by arrows.