Figure 6
From: Intrinsically active MEK variants are differentially regulated by proteinases and phosphatases
Stability and intrinsic activity of MEKs and their mutants. (A) Effect of translation and protease inhibitors on MEKs and their mutants. Cos7 cells were transfected with the indicated constructs, and 24 hours later the cells were treated with either Cyclohexamide (C; 1 μg/ml) or MG132 (M, 10 μM) for 8 h or left untreated. Then the cells were harvested with sonication (2 × 7 sec) and subjected to western blotting with anti gMEK and gERK as indicated. Quantification of the results appears in the lower panel. The bargraph represent the average and standard error of 2or 3 experiments *P < 0.05 by student’s t-test. (B) Effect of Cyclohexamide on the expression of WT-MEK1, Q56P-MEK1 and Y130C-MEK1. COS7 cells were transfected with WT-MEK1, Q56P MEK1 and Y130C-MEK1. Twenty-four hours later, the cells were treated with Cyclohexamide (CHX; 1 µg/ml) for the indicated times (bottom). The cells were then harvest and subjected to western blotting using anti gMEK and gERK Abs. The half-life of the GFP-MEK proteins is indicated in the right.
