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. 2010 Apr 16;285(24):18283–18290. doi: 10.1074/jbc.M109.079137

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 6. — Activation of PAK1 in Lkb1(+/−) mouse HCCs and Lkb1(−/−) MEFs. A, activation of PAK1 in Lkb1(+/−) mice HCCs. Cellular levels of phospho-PAK1, PAK1, and β-actin in tissue lysates were determined by Western blotting with anti-phospho-PAK1 (Ser¹⁴⁴), anti-phospho-PAK1 (Ser^199/204), anti-PAK1, and anti-β-actin. Two wild-type livers (W1 and W2), pairs of tumor (H1 and H2), precancerous lesions (F1 and F2) and adjacent normal tissue (N1 and N2) from two Lkb1(+/−) mice were analyzed. B, increased PAK1 activity in Lkb1(−/−) MEFs. Cellular levels of phospho-PAK1, PAK1, and β-actin in tissue lysates were determined by Western blotting with the indicated antibodies. For in vitro kinase assay, cell lysates were prepared from MEF cells and immunoprecipitated using an anti-PAK1 antibody. The precipitates were used for in vitro kinase assays using GST-VASP-(158–277) as a substrate. The phosphorylation of GST-VASP was visualized using BAS-5000 Bio-imaging Analyzer. Similar results were obtained in three independent experiments.