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. 2011 Jun 24;6(6):e21482. doi: 10.1371/journal.pone.0021482

Figure 9. ERK resides upstream of FAK in ECSM2-regulated FGFR-ERK-FAK pathway.

Figure 9

(A) Blockade of ERK activation eliminates the upregulation of pFAK(Y397) and the downregulation of pFAK(S910) (bFGF-induced) in MS1 cells expressing ECSM2-GFP versus GFP control cells. Serum-starved cells were pretreated with DMSO (control) or PD98059 (50 µM) for 1 h prior to stimulation with vehicle (-) or bFGF (10 ng/ml) for 15 min. Protein extracts were analyzed by immunoblotting with anti-pERK, anti-total ERK, anti-pFAK(S910), anti-pFAK(Y397), or anti-total FAK, as indicated. (B and C) Statistical analysis (densitometry) of pooled data of pFAK(S910) and pFAK(Y397) from three independent experiments. Data are mean±SEM. **, P<0.01; ns, not statistically significant. (D) Effect of inhibition of ERK activity on bFGF-induced cell migration. Serum-starved MS1 cells stably expressing GFP or ECSM2-GFP were pretreated with DMSO (control) or PD98059 (50 µM) for 1 h prior to transwell assays, as detailed in Methods. The cells were allowed to migrate across the membrane of the transwell insert in response to bFGF (10 ng/ml) in serum-free media for 5 h. The number of migrated cells per imaging field for each condition was counted. Data are mean±SEM (n = 12). **, P<0.01; ns, not statistically significant.