Figure 2.

Pitstop2 treatment inhibits HGF-stimulated Akt phosphorylation. RPE cells were treated with 10 μM pitstop2 or vehicle control (0.1% DMSO) for 30 min. Following drug treatment, cells were stimulated with either 5 ng/mL EGF or 50 ng/mL HGF for 5 minutes in the continued presence or absence of the inhibitor, or left unstimulated (basal). Whole-cell lysates were prepared, resolved by immunoblotting, and probed with anti-phospho-Akt (pS473) anti-total-Akt or anti-actin antibodies. All methods were performed as described previously.⁵ HGF was obtained from Life Technologies (Carlsbad, CA). Shown in the left panel are representative immunoblots, and in the right panel the mean ± SE of the pAkt values in each condition (n = 4, *, p < 0.05).