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. 1999 Nov 23;96(24):14019–14024. doi: 10.1073/pnas.96.24.14019

Figure 2.

Figure 2

FTS interrupts ERK activity in human melanoma cells. Human 518A2/N-Ras (L61) melanoma cells (A) and a vector-transfected control (B and C) were grown for 24 h in DMEM/10% FCS in the presence of 0.1% DMSO (zero drug control) or in the presence of the indicated concentrations of FTS. ERK activity (A and B) was then determined in cell lysates with MBP as a substrate as described in Materials and Methods. The phosphorylated MBP bands, visualized by autoradiography, demonstrate the dose-dependent decrease in ERK activity. Densitometric analysis (see Materials and Methods) of data obtained from representative experiments indicated a decrease in ERK activity of 25–32%, 39–58%, and 65–95%, respectively, in the presence of 5, 10, and 25 μM FTS either in the 518A2/N-Ras (L61) or the vector-transfected control cells. The total amount of Raf-1, MEK, ERK remain unaltered in 518A2 cells treated 24 h with FTS (C). Raf-1, MEK, and ERK in cell lysates were determined by immunoblotting with the corresponding Abs.