Fig. 5.

Glycosaminoglycan chains do not affect tPA-catalyzed plasmin generation. (A) Western blot analysis of NG2 secretion by wt and xylotransferase^−/− CHO cells. Conditioned media (CM) from wt CHO cells was treated with chondroitinase to remove GAG chains before the western blot analysis, whereas the xylo^−/− CHO CM was electrophoresed without prior chondroitinase treatment. Chondroitinase-treated recombinant purified NG2 was electrophoresed in parallel as a loading control. (B) wt CHO cells and xylo^−/− CHO cells were immunostained for membrane-bound NG2 (red). Nuclei were stained using DAPI (blue). (C) wt CHO CM and xylotransferase^−/− CM were scored for their ability to enhance plasmin generation in the presence or absence of tPA and plg. Nonconditioned medium was used as a negative control. (D) Binding assays with wt cells or Xylo^−/− CHO cells in the presence or absence of tPA. Error bars represent S.E.M where **P < 0.001, #P < 0.05 by ANOVA followed by Bonferroni’s post hoc test. [Color figure can be viewed in the online issue, which is available at www.interscience.wiley.com.]