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. Author manuscript; available in PMC: 2009 Apr 1.
Published in final edited form as: Cell Signal. 2007 Dec 8;20(4):675–683. doi: 10.1016/j.cellsig.2007.12.003

Figure 7. MLK-3-, but not MEKK1-mediated phosphorylation of JNK by SB202190 or SB203580.

Figure 7

Figure 7

(A) A549 cells were transfected with either wildtype MEKK1 or kinase-inactive MEKK1 (dnMEKK1) expression vectors using the transfection protocol as described in the Methods section. After 48 hours, these cells were treated with either SB202190 or SB203580 (2 µM each) for 2 hours. Cell lysates were prepared and phosphoJNK was detected, as described. (B) MLK-3 is required for the phosphorylation of JNK due to treatment of cells with SB202190 or SB203580. A549 cells were either transfected with NT siRNA or with MLK-3 siRNA as described in the Methods section. After 48 hours, these cells were treated with either SB202190 or with SB203580 (1µM) for 2 hours. Cell lysates were prepared and phospho-JNK (top panel), unphosphorylated JNK (second panel from top), MLK-3 (third panel from top), and β-actin (fourth panel from top) were detected using respective antibodies, as described in the Methods section.