Figure 5

E2-induced changes in FAS and FGFR2 proteins. A) MCF-7p and MCF-7AKT cells express FAS proteins of different mobility with or without E2 treatment. Cells were treated with E2 for indicated time (in hours) and cell lysates were subjected to western blotting. B) Dominant-negative AKT (KD-AKT) inhibits E2-induced alternative splicing of FAS as evident from qRT-PCR assays. *p<0.03, total vs isoform at 3 and 6 hour E2 treated condition in control cells. C) The effect of E2 on FGFR2 protein in MCF-7p and MCF-7AKT cells. *p=0.0004 untreated vs six hour E2 treated MCF-7p cells (for faster migrating FGFR2). *p=0.03 untreated MCF-7p versus untreated MCF-7AKT cells. Two exposures of FGFR2 blot are shown. D) ERα is essential for E2 regulated FGFR2 expression. Cells were treated with control luciferase siRNA (Luci) or ERα siRNA. After three days of siRNA transfection, cells were additionally treated with ethanol or E2 for 24 hours. Two exposures of FGFR2 and ERα blots are shown.