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. Author manuscript; available in PMC: 2013 Jul 25.
Published in final edited form as: Exp Cell Res. 2010 Jul 22;316(17):2825–2832. doi: 10.1016/j.yexcr.2010.07.012

Figure 1. The translation initiation factor eIF4E regulates RhoA protein levels in breast tumor cells.

Figure 1

A. MDA-MB-231 breast tumor cells were transfected transiently with a flag-tagged eIF4E-expressing or control vector. Equivalent amounts of protein from these cells were subjected to SDS-PAGE, and immunoblotted with antibodies specific for RhoA, the flag tag, or β–actin, followed by the appropriate species secondary antibody. B. MDA-MB-231 cells were transfected transiently with an eIF4E-specific or control siRNA. Equivalent amounts of protein from these cells were subjected to SDS-PAGE and immunoblotted with antibodies specific for RhoA, eIF4E, or β–actin, followed by the appropriate species secondary antibody. C. MDA-MB-231 cells were incubated in medium containing Rapamycin at the indicated concentration for 4 hours. Equivalent amounts of protein from these cells were subjected to SDS-PAGE and immunoblotted with antibodies specific for RhoA, phospho(Ser65)-eIF4E binding protein (p-4E-BP1), representing the inactive form of the the eIF4E inhibitor eIF4E-BP1, eIF4E-BP1, or β–actin, followed by the appropriate species secondary antibody. For A-C, relative RhoA expression, as well as the ratio of phospho-eIF4E-BP1 to total cellular eIF4E-BP1, was quantified by densitometry using ImageJ software (NIH). Similar results were observed in at least 3 independent trials.