Fig. 2.

Epac activates syndecan-2 translocation into rafts in SK-Mel-2. A: immunocytochemistry for syndecan-2 (red) and FLAER (green) is shown. Top, cells were incubated in the presence or absence of 50 μM 8-pMeOPT for 15 min. 8-pMeOPT induced colocalization of syndecan-2 with lipid rafts (yellow). Bottom, cells overexpressing LacZ or Epac1 were incubated in the presence or absence of 10 μg/ml CXD. Epac1 overexpression increased colocalization of syndecan-2 with lipid rafts (yellow). CXD decreased such colocalization. B: immunoblot showed that syndecan-2 expression was not changed by Epac1 overexpression. n = 4. C: immunoblot showed that Epac1 overexpression increased syndecan-2 expression in rafts-rich fraction. Flotillin was used for a marker of lipid rafts. n = 4. D: migration assay showed that CXD (10 μg/ml) decreased basal and Epac1 overexpression-induced cell migration. Syndecan-2 siRNA also inhibited Epac1 overexpression-induced cell migration. n = 4. E: immunoblot showed decreased syndecan-2 expression with siRNA. n = 4.