Fig 5.

Effects of commercially available PI3K/Akt inhibitors and BEZ-235 on LCMV MG-derived reporter gene expression. (A) Drug effects on MG-derived reporter gene expression. 293T cells were transfected with pC-T7, pMG-CAT, pC-NP, and pC-L as described previously (18, 19, 29, 31, 32). After 5 h of transfection, medium was replaced with fresh medium containing Akt-IV (0.5 μM), Akt-VIII (2.0 μM), LY (20 or 50 μM), or BEZ-235 (0.05, 0.5, or 5 μM). At 24 h posttransfection, cell lysates were prepared for CAT ELISA. CAT expression levels from vehicle (DMSO)-treated cells were set to 1.0 to normalize CAT expression levels from the other samples. (B) Drug effects on Pol-II-based transcription. 293T cells (8 × 10⁴/96-well plate) were transfected with pC-Fluc using Lipofectamine 2000 (Invitrogen), and 12 h later, medium was replaced with fresh medium containing the indicated compounds at the indicated concentrations. At 24 h post-compound treatment, levels of Fluc were determined using the Steady Glo assay (Promega) and a luminometer (Centro LB 960; Berthold Technologies). The viability of DMSO-treated control cells was set at 1.0. Data are averages and standard deviations from three independent experiments.