Figure 4. Kinase Assay validates FTS hits as inhibitors.

MAP2K4 kinase assays were performed as described in Methods using either p38 MAPK (K53A) or JNK1 (K55M) as substrates, and was done using a range of concentrations of either staurosporine A or daidzein B. Representative phosphorylated JNK1 and phosphorylated p38 MAPK and total JNK and p38 MAPK Western blots are depicted. C. A plot of densitometric measurement of bands from separate experiments utilizing Staurosporine (2X each on p38 and JNK1 substrates) is depicted with intensity measurements shown as mean ± S.D along with the curve fit used for determining the IC₅₀ values; Red – JNK1 and Black – p38MAPK. D. 3 indicated concentrations of either MAP2K4-EE or MAP2K4-AC were employed in the kinase assay and the western blot performed as before.