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Figure 8


Fig. 8. Activation of Rho GTPases upon fibroblast adhesion to collagen I. Wild-type (WT) and {alpha}2-deficient (-/-) fibroblasts were serum-starved for 24 hours and subsequently seeded at the same cell density on collagen I. After 60 to 120 minutes of adhesion the cells were lysed and active, GTP-bound RhoA, Rac1 and Cdc42 were pulled down as described in the Materials and Methods. (A) Representative blots of total (lysate) and GTP-bound RhoA, Rac1 and Cdc42 (pull-down) after 90 (lane 1) and 120 (lane 2) minutes of adhesion to collagen for wild-type (WT) and {alpha}2-deficient (-/-) fibroblasts. (B) Band intensity of total and GTP-bound proteins were measured by densitometry on blots from different experiments and the relative amounts of active, GTP-bound proteins were normalized to those of the corresponding lysates (arbitrary units). The graphs show the results from different experiments performed for 60, 90 and 120 minutes as indicated below each set of columns. White columns, control fibroblasts; black columns, {alpha}2-deficient fibroblasts.





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