Fig. 2.

Src family inhibitors suppress PACAP-induced ERK phosphorylation. A: representative Western blot analysis. PAC1 receptor cell cultures were treated with PACAP (P27, 25 nM) alone for 15 min. Other cell cultures were pretreated with Src family inhibitors PP2 (10 µM) or Src-1 (2 µM), or with the inactive PP2 analog PP3 (10 µM) for 15 min before exposure to PACAP in the presence of Src compounds for an additional 15 min. B: PACAP stimulated ERK activation ~7-fold at 15 min. Following pretreatment with PP2 or Src-1, the PACAP-induced ERK phosphorylation was significantly reduced >50%. In contrast, pretreatment with PP3 did not significantly reduce the PACAP-induced ERK phosphorylation. All data were normalized to total ERK levels in the same samples. Data represent mean fold change ± SE; n = 3 independent experiments. *Different from CTL. +Different from P27 following pretreatment with PP2 or Src-1; all at P < 0.05.