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. 2010 Dec 1;286(6):4199–4208. doi: 10.1074/jbc.M110.155093

FIGURE 1.

FIGURE 1.

The expression of C4ST-1 gene was suppressed in L-Wnt-3a cells. A, the disaccharide composition of HS chains produced in L and L-Wnt-3a cells was analyzed. GAG chains isolated from L and L-Wnt-3a cells were digested with a mixture of heparitinase and heparinase, labeled with the fluorophore 2-aminobenzamide, and analyzed by HPLC. The percentage of different HS disaccharides is shown. Results are the means ± S.E. for two experiments. ΔDiHS-0S, ΔHexUAα1–4GlcNAc; ΔDiHS-6S, ΔHexUAα1–4GlcNAc(6-O-sulfate); ΔDiHS-NS, ΔHexUAα1–4GlcN(2-N-sulfate); ΔDiHS-diS1, ΔHexUAα1–4GlcN(2-N-sulfate,6-O-sulfate); ΔDiHS-diS2, ΔHexUA(2-O-sulfate)α1–4GlcN(2-N-sulfate); ΔDiHS-triS, ΔHexUA(2-O-sulfate)α1–4GlcN(2-N-sulfate,6-O-sulfate). B, NDST1 (AF049894), HS2ST1 (AF060178), and HS6ST1 (NM_015818.2) mRNA expression levels in L (open bars) and L-Wnt-3a cells (closed bars) were measured using quantitative real-time PCR. Transcript levels of each gene are expressed relative to GAPDH mRNA levels ± S.D. Two independent experiments were run for each data set. C, the disaccharide composition of CS chains produced in L and L-Wnt-3a cells was analyzed as described in A. The percentage of different CS disaccharides is shown. ΔDi-0S, ΔHexUAα1–3GalNAc; ΔDi-6S, ΔHexUAα1–3GalNAc(6-O-sulfate); ΔDi-4S, ΔHexUAα1–3GalNAc(4-O-sulfate); ΔDi-diSD, ΔHexUA(2-O-sulfate)α1–3GalNAc(6-O-sulfate); ΔDi-diSE, ΔHexUAα1–3GalNAc(4-O-sulfate,6-O-sulfate); ΔDi-triS, ΔHexUA(2-O-sulfate)α1–3GalNAc(4-O-sulfate,6-O-sulfate). D, C4ST-1 (NM_021439.2), C4ST-2 (NM_021528), D4ST (BC043700), and GalNAc4S-6ST (NM_029935.5) mRNA expression levels in L (open bars) and L-Wnt-3a cells (closed bars) were analyzed using quantitative real-time PCR as described in B. E, poly(A)+ RNA was isolated from L and L-Wnt-3a cells and analyzed by Northern blot hybridization using DNA probes specific for C4ST-1 and GAPDH.