FIGURE 2.

Butaprost induced Src and EGFR activation. A, butaprost dose-dependently induced phosphorylation of Src and EGFR. Mice were treated with butaprost (50 or 100 nmol) and sacrificed at 2 h after treatment. *, p < 0.05 versus vehicle-treated mice. IP, immunoprecipitation; WB, Western blot. B, butaprost (100 nmol) induced phosphorylation of Src and EGFR in a time-dependent manner. C, AG1478 and PP2 decreased EGFR phosphorylation. The dorsal skin was treated with PP2 (100 nmol) or AG1478 (100 nmol) for 30 min before butaprost (100 nmol) treatment, and skin sections were isolated 2 h later. *, p < 0.05 versus butaprost-treated mice. In A–C, Src and EGFR were immunoprecipitated from skin lysates (200 μg) by an anti-Src or an anti-EGFR antibody. p-Src and p-EGFR were detected by Western blot analysis by using a monoclonal anti-p-Tyr antibody. The data are representative of two independent experiments. In A and B, skin lysates (50 μg) were immunoblotted for p-Src (Tyr⁴¹⁶). In A–C, the intensities of the bands were determined by densitometry as described in the legend for Fig. 1. In A and C, the relative intensities are presented as mean value ± S.D. (n = 3) from two independent experiments.