Effect of LN on FAK phosphorylation and its downstream signaling kinases (Akt and ERK1/2) activity in AsPC-1 cells. A, AsPC-1 cells were treated as described in results for the indicated time and then subjected to western blot analysis to detect the expression of pFAK (pY397), p-Akt (pS473), p-ERK1/2 and their total proteins. The membranes were probed with anti-β-actin antibody to ensure even loading of proteins in each lane. B, For semiquantitation, the intensities of bands for each cell line were calculated using a densitometer. Bars are mean ± SE of relative pFAK/FAK, pAkt/Akt or pERK/ERK ratio (normalized to that of 0 min, which was given a value of 1) for three independent experiments.