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. Author manuscript; available in PMC: 2012 Dec 1.
Published in final edited form as: Cell Signal. 2011 Jul 23;23(12):1978–1987. doi: 10.1016/j.cellsig.2011.07.008

Figure 4. Overexpression of SPARC in Daoy medulloblastoma cells inhibits Rho-Rac-Cdc42 GTPase activity.

Figure 4

(A) Daoy-P, Daoy-EV, and Daoy-SP cells were grown to 70% confluence and the cell lysates were either incubated with the GST-TRBD fusion beads for 60 minutes or used for SDS-PAGE analysis. The lysates incubated with beads were then washed three times, resuspended in SDS loading buffer, and boiled for 10 min. SDS-PAGE and Western blot analyses were performed. (A) Western blot analysis for Active RhoA, total RhoA, active Rac1, total rac1, active Cdc42, total Cdc42, cofilin and pcofilin. (B) Densitometric analyses of western blots described in A. Results are expressed as the ratio between active and total levels of RhoA, Rac1, and cdc42. Phosphorylation is calculated as the ratio of the phosphorylated versus nonphosphorylated forms. Data are representative of three independent experiments (p<0.05).