Skip to main content
. 2011 Sep 6;105(7):953–960. doi: 10.1038/bjc.2011.349

Figure 5.

Figure 5

The E2 mediated signal through ERα, c-Src, PI3K and mTOR in breast cancer lines. (A) MCF-7 and T47D cells were cultured in steroid-free medium for 48 h. The ERα inhibitor ICI 182780 (10 μM), c-Src inhibitor PP1 (10 μM) and PI3K inhibitor LY294002 (20 μM) were added for 30 min before the stimulation with 10 nM E2. The cells were incubated in the presence and absence of 10 nM E2 for 15 min. At the end of time point, cells were harvested and immunoblotting was used to detect the phosphorylation of Akt on Ser473, mTOR on Ser 2448 and the levels of Akt and mTOR. (B) Steroid-deprived MCF-7 and T47D cells were treated with ERα, c-Src, PI3K and mTOR (rapamycin 50 nM) inhibitors for 30 min, followed by 10 nM E2 for 30 min. Cells were lysed and immunoblotted with phosphorylated specific and total form of p70S6K and 4EB-P1 antibodies. (C) Cells were transfected with siRNA to c-Src, Akt and mTOR, followed by steroid starvation for 48 h. Cells were stimulated with E2 for 30 min and lysed. Cell lysates were subjected to immunoblotted with phosphorylated specific and total form of p70S6K and 4EB-P1 antibodies.