Figure 5. Fluorophore-assisted carbohydrate electrophoresis (FACE) analysis of keratan sulfate secreted by keratocytes. Keratan sulfate from culture medium was digested with keratanase II and endo-β-galactosidase, and the fragments were analyzed by fluorophore-assisted carbohydrate electrophoresis (FACE). A: Marked bands show products previously determined to be derived from bovine keratocyte keratan sulfate: 0S is the unsulfated disaccharide generated by endo-β-galactosidase; 1S is the monosulfated disaccharides generated by keratanase and endo-β-galactosidase; Gal-S is galactose sulfate, a product of keratanase digestion; and 2S is disulfated disaccharide, a product of keratanase digestion. Unmarked bands represent non-specific and/or unidentified products present in the dye or the digest. Keratan sulfate samples were isolated from the conditioned media of attached (AT) and spheroid (SP) cells cultured five days in Advanced-DMEM with (+) or without (-) TGFβ1 at 2 ng/ml. B: The chart shows the quantification of the keratan sulfate products in gels similar to that shown in A. Blue bars (left two of each group) are attached cells whereas red/pink (right two of each group) are from sphere cultures. The darker bars (Bars 1 and 3 of each group) are from cultures without TGFβ and the lighter bars (Bars 2 and 4 of each group) were treated with TGFβ. Error bars show standard deviation of triplicate analyses.