Fig. 2. Activation of PI-3K and MAPK by IGF-1 and EGF. Western blot with phosphorylation-specific antibodies against MAPK (A, top; B, top) or Akt (B, bottom) showing the effect of growth factor treatment and pharmacological inhibitors on activation of MAPK and Akt. The lower panel in A and the middle panel in B show total (phosphorylated and unphosphorylated) MAPK (ERK2) as loading control. (A) Time course of MAPK activation after stimulation with EGF in single keratinocytes treated with vehicle control (DMSO) or 10 µM PD98059. Keratinocytes were stimulated for the indicated times. (B) Activation of MAPK and Akt in preconfluent cultures of keratinocytes. Cells were pretreated with DMSO or the inhibitors PD98059 (10 µM) and LY294002 (25 µM), respectively, and then stimulated with growth factors for 15 minutes.

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