FIGURE 3.

Disaccharide compositions of GAGs from WT, HS6ST-1^−/−, HS6ST-2^+/−, HS6ST-2^−/−, and HS6ST-1^−/−/HS6ST-2^−/−-FSMCs. FSMCs were prepared from WT, HS6ST-1^−/−, HS6ST-2^+/−, HS6ST-2^−/−, and HS6ST-1^−/−/HS6ST-2^−/− embryos as described under “Experimental Procedures.” HS/heparin and chondroitin sulfate were isolated from FSMCs, digested with a mixture of heparitinase-I, heparitinase-II, and heparinase, and chondroitinase ABC, and subjected to reverse phase ion pair chromatography as described under “Experimental Procedures.” The histograms show the percentage compositions of the unsaturated disaccharides in the HS/heparin (A and B) and in chondroitin sulfate (C) isolated from the FSMCs of WT (open bar), HS6ST-1^−/− (Crossed hatched bar), HS6ST-2^+/− (gray bar), HS6ST-2^−/− (hatched bar), and HS6ST-1^−/−/HS6ST-2^−/− (filled bar). A and B, the values (heights of the histograms) shown are the means of triplicate experiments, and the bars represent the standard deviation. ΔDi-0S, ΔDi-NS, ΔDi-6S, ΔDi-(N,6)diS, ΔDi-(N,2)diS, and ΔDi-(N,6,2)triS are the same as described in the legend for Fig. 1A. C, ΔDi-0S, ΔDi-4S, ΔDi-6S, ΔDi-diS_E, and ΔDi-diS_D are unsaturated disaccharides derived from -HexA-GalNAc-, -HexA-GalNAc(4SO₄)-, -HexAGalNAc(6SO₄)-, -HexA-GalNAc(4,6)SO₄-, and -HexA(2SO₄)-GalNAc(6SO₄)- units of the GAGs, respectively.