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Activated Pak1 induces MLC phosphorylation in the absence of MAPK activation. Control NIH-3T3 cells or NIH-3T3 cells expressing various forms of Pak1 were starved overnight in media containing 0.5% CS, then treated with the indicated compounds (PDGF, 10 ng/ml or LPA 5 μg/ml) for 10 min. Immunoblots of cell lysates were probed with anti-HA (to detect transgene expression), and antibodies against the activated (phosphorylated) forms of Jnk, Erk, and MLC.
