FIGURE 7.

Fe65 potentiates the estrogen stimulation of ERα target gene expression in BCa cells. A, MCF-7 cells were transfected with empty vector (Ctrl), Fe65, or PTB2 deletion mutant and treated with EtOH or 10⁻⁸ m E2 for 2 h. Cellular extracts were subjected to IB analyses with the indicated antibodies. The expression levels of cyclin D1 and c-Myc were quantified with ImageJ and normalized with cognate β-actin signals. The normalized signals were divided by the signal of control cells that were transfected with empty vector or scrambled siRNA and treated with EtOH and are shown at the bottom of the blots. B, MCF-7 cells stably transfected with empty vector or Fe65 were treated, and levels of protein expression determined as in panel A. C and D, T47D (panel C) and BT474 (panel D) cells transiently transfected with siCrtl or siFe65 were treated, and levels of protein expression determined as in panel A. E, MCF-7 cells stably transfected with empty vector or Fe65 were treated as in panel A but for 1 h. Total RNA was extracted and subjected to real time RT-PCR analyses. The levels of c-Myc mRNA were normalized with GAPDH and are expressed as -fold of the EtOH control. F, T47D cells transiently transfected with siCrtl or siFe65 were treated as in panel A but for 1 h. Total RNA was extracted and subjected to real time RT-PCR. The levels of cyclin D1 and c-Myc mRNA were normalized with GAPDH and are expressed as -fold of the EtOH controls.