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BIX 02189 is a selective MEK5/ERK5 inhibitor with an IC50 of 59 nM. BIX02189 was reported to inhibit catalytic function of purified, MEK5 enzyme. BIX02189 blocked phosphorylation of ERK5, without affecting phosphorylation of ERK1/2 in sorbitol-stimulated HeLa cells. BIX02189 also inhibited transcriptional activation of MEF2C, a downstream substrate of the MEK5/ERK5 signaling cascade, in a cellular trans-reporter assay system. BIX02189 may offer novel pharmacological tools to better characterize the role of the MEK5/ERK5 pathway in various biological systems.