FIGURE 7.

Versican overexpression induces transition of normal human dermal fibroblasts to a myofibroblast phenotype. A, Western blotting of cell monolayers with an antibody to versican (anti-GAGβ) shows that increased versican V1 isoform expression (arrow) was achieved by transient transfection. Versican migrates as an ∼350-kDa species when deglycosylated with chondroitinase ABC (arrow, upper panel) Coincident with this, there were increased levels of DPEAAE and pSMAD2 immunoreactivity (representative of n = 2). B, Vcan-overexpressing cells had increased levels of SMA (center panel) (representative of n = 3). C, quantification of SMA by densitometry of Western blots shows expression of significantly higher levels in normal human dermal fibroblasts by Vcan overexpression (*, p < 0.01, n = 3). D, collagen gel contraction assay showed that Vcan-overexpressing cells can contract a collagen gel to a greater extent. E, quantification of collagen gel contraction assay shows that V1 versican-overexpressing cells are contractile to a greater extent than control vector-transfected cells (*, p < 0.01, n = 3 independent transfections with three gels per experiment). Error bars indicate S.D.