Figure 2.

Activation of EGFR pathways in both in vitro and in vivo models of peritoneal metastasis. The statuses of EGFR signaling molecules (EGFR, Akt and ERK) were examined in ES-2 cells in spheroid cultures and in ascites using an immunoblotting analysis. Phosphorylation status was examined to evaluate the activated forms of these proteins. The analyzed sites of phosphorylation on EGFR, Akt and ERK were Tyr¹⁰⁶⁹, Ser⁴⁷³ and Thr²⁰²/Tyr²⁰⁴, respectively. (A) ES-2 cells were seeded into a spheroid culture plate on day 1 and were analyzed on days 4 and 7. Ctrl: monolayer culture. (B) ES-2 cell aggregates in ascites were collected from mice with peritoneal metastasis (n=6, mouse ID#1-6)