Figure 3.

Rapamycin inhibited mTOR activity and reduced BMI-1 and cyclin E1. A. Rapamycin inhibited mTOR pathway and reduced BMI-1 and cyclin E1 protein. Y79 cells were treated with the indicated concentration of rapamycin for 48 h. The indicated protein levels were determined by Western blot analysis as described in “Materials and methods”. β-Actin protein levels were used as a loading control. All these experiments were replicated at least thrice, and a representative experiment was shown in each panel. B. Rapamycin decreased the mRNA expression of Bmi-1. Y79 cells were treated with the indicated concentration of rapamycin for 48 h. The indicated mRNA levels were determined by semi-quantitative RT-PCR analysis as described in “Materials and methods”. β-Actin mRNA levels were used as a loading control. All these experiments were replicated at least thrice, and a representative experiment was presented in each panel.