Fig. 8. Erk1/2 and Akt phosphorylation in PDGF-BB-stimulated mesonephric cells: inhibition by U0126 and Ly294002. Quiescent cells were incubated with U0126 or Ly294002 and then stimulated with different concentrations of PDGF-BB for 15 minutes or overnight, as described in Materials and Methods. Lysed cells were separated by SDS-PAGE and blotted filters were incubated with (A) antibody for phospho-Erk1/2 (upper panel) and for nonphosphorylated forms as control (lower panel); or (B) antibody for phospho-Akt (upper panel) and for nonphosphorylated form as control (lower panel).