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Thrombin stimulates Pak1 activity in SUM149 cells. Cell lysates were made from SUM149 and MDA-MB-231 cells treated with thrombin (0.5 U/mL) for various times. After immunoprecipitation of the cell lysates with anti-Pak1 antibody or IgG, Pak1 kinase activity was analyzed by kinase assay using MBP as a substrate. The middle panels are Ponceau S-stained blots showing MBP. The lower panels are the total Pak1 level determined by immunoblotting using anti-Pak1 antibody.
