Combined mTOR and MEK inhibition attenuates cellular proliferation and increases the apoptotic response.

<p>(<b>A</b>) The indicated cells were treated for 24 hrs. with rapamycin or BEZ235 and immunoblotted with the indicated antibodies. (<b>B</b>) 786-0 and RCC4 cells were treated increasing doses of GSK212 for 24 hrs. and immunoblotted with the indicated antibodies. (<b>C</b>) 786-0 and RCC4 cells were treated for 24 hrs with rapamycin and BEZ235 in the presence of Edu. Edu incorporation was assessed by flow cytometry. (<b>D</b>) 786-0 and RCC4 cells were treated with the indicated compounds for 24 hrs. Whole cell extracts were immunoblotted for the cell cycle related proteins indicated. (<b>E</b>) 786-0 and RCC4 cells were treated with indicated drugs and assessed for viability on day 4 using CellTiter-Glo 4. (<b>F</b>) 786-0 and RCC4 cells were plated, allowed to attach, and treated with the indicated drug(s). Photographs of wells containing 786-0 (day 11) and RCC4 (day 17) cells fixed with 4% PFA and stained with 0.1% crystal violet. (<b>G</b>) 786-0 and RCC4 cells were treated with the indicated compounds for 24 hrs. Whole cell extracts were immunoblotted with the indicated antibodies.