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. 2008 Jan 2;28(5):1528–1540. doi: 10.1128/MCB.02061-07

FIG. 3.

FIG. 3.

Transfection of Snail1 to RWP-1 cells up-regulates Akt activity and decreases PTEN mRNA levels. Control (cont) RWP-1 cells stably transfected with pcDNA3 or with pcDNA3-Snail-HA were irradiated and analyzed. (A) Apoptosis was determined as described previously for nonirradiated cells (−) or for cells 24 h after γ radiation (+). The figure shows the averages ± ranges of three experiments performed. (B) Total cell extracts were prepared from nonirradiated cells (−) or 24 h after γ radiation (+) and analyzed by Western blotting (WB) with MAb against HA, PThr308-Akt, caspase-3 (Casp-3) or annexin 2 as a loading control. (C) mRNA was prepared from RWP-1 cells at the indicated time points, and levels of PTEN mRNA were detected by quantitative RT-PCR. Results are presented as the averages ± SDs (error bars) of three independent experiments. When we compared the levels of PTEN mRNA between control and Snail1-expressing cells at the same time points, the differences were significant at all time points with a P value of <0.01.