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Figure 4


Fig. 4. Reactive astrocytes produce more 4-sulfated than 6-sulfated chondroitin sulfate GAG chains. (A) Conditioned medium from 3 day culture was concentrated and digested with cABC, followed by immunoblot analysis. (B) Conditioned medium collected at indicated time was treated with cABC and subjected to ELISA without concentration. Monoclonal antibodies 2B6/MAB2030 and 3B3/MAB2035 were used against 4-sulfated and 6-sulfated GAG chains, respectively. (C) C4ST1 mRNA was rapidly increased upon induction of reactive astrocytes by TGFβ1. RNA was prepared from astrocytes at indicated time after TGFβ1 treatment and quantitative RT-PCR was performed. Open column, control; filled column, TGFβ1. (D) Transcript of the gene encoding C4ST1 but not C6ST1 was upregulated 3 days after TGFβ1 treatment of astrocytes. Open column, control; filled column, TGFβ1. (E) Upregulation of C4ST1 protein was detected with chicken anti-C4ST1 peptide antibody 3 days after TGFβ1 treatment. Lysates from HeLa cells transfected with pTracer-C4ST1 were used as a positive control.





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