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Fig. 10. Inhibition of PI 3-kinase (LY 294002) and MEK1 (PD 98059) prevents HGF/SF-induced phosphorylation of PKC-{zeta}. Western blot analyses of HaCaT cells that were left untreated (media change) or were exposed to HGF/SF (for 10 minutes at 100 ng/ml) after preincubation with (A) PI 3-kinase inhibitor LY 294002 (LY; at 10 µM for 60 minutes), with (B) MEK1 inhibitor PD 98059 (PD; at 50 µM for 60 minutes) or with solvent only (DMSO, 0.1%). Cellular extracts were immunoprecipitated (IP) by specific anti-PKC-{zeta} antibody (Santa Cruz) prior to immunoblotting (IB). Immunoblotting with pan-PKC-{zeta} antibody (upper panel) and with phospho-PKC-{zeta} antibody (lower panel). Immunoblots displayed are representative of two that were performed revealing comparable results.