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Figure 6


Fig. 6. Depletion of C6ST1 by siRNA does not alter the production of 4-sulfated chondroitin sulfate GAG chains nor repellent activity in reactive astrocytes. Astrocytes transfected with C6ST1 siRNA were treated with or without TGFβ1 as in Fig. 5, and RNA and conditioned medium were prepared. Quantitative RT-PCR for (A) C4ST1, (B) C6ST1. ELISA to measure the amount of (C) 4-sulfated GAG chains with MAB2030, (D) 6-sulfated GAG chains with MAB2035. (E) Axonal guidance spot assay was performed (*P<0.01 compared with scrambled siRNA without TGFβ1; ANOVA). Although C6ST1 siRNA reduced C6ST1 mRNA and production of 6-sulfated GAG chains, induction of C4ST1 mRNA and 4-sulfated GAG chains by TGFβ1 were unaffected, and repellent activity of TGFβ1-treated conditioned medium was observed.





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