Fig. 2.

PMA induces expression of Renilla luciferase from the pRL-TK plasmid in HEK293 cells. Cells were transfected with the pRL-TK plasmid and treatment as indicated was added at 24 hours post-transfection. The cells were then harvested at the appropriate times and Renilla luciferase assays were carried out. A and B: HEK293 cells transfected with the pRL-TK plasmid were left non-treated or treated as shown for 8 hours; ionomycin was used at 1 μM.; ethanol and DMSO were added to a final concentration of 0.1% (v/v). Fold change was calculated by setting the normalized Renilla luciferase relative luminescence units (RLU) for the non-treated sample as 1. The results shown are the mean and SD of three independent experiments. C: HEK293 and HeLa cells (HEK293* indicates HEK293 cells obtained from another laboratory) transfected with the pRL-TK plasmid were left non-treated or treated with PMA for 8 hours. Fold change was calculated by setting the normalized Renilla luciferase RLU for the non-treated sample of the corresponding cell line as 1. The results shown are the mean and SD of three independent experiments. D: HEK293 cells transfected with the pRL-TK plasmid together with a GFP-expressing plasmid and an empty vector plasmid (either pcDNA3 or pUC19) as shown were left non-treated or treated with PMA for 8 hours. Fold change was calculated by setting the normalized Renilla luciferase RLU for the non-treated sample transfected with pGFP/pcDNA3 as 1. The results shown are the mean and SD of three independent experiments. E: HEK293 cells transfected with the pRL-TK plasmid were treated with PMA for various durations of time as shown. The results shown are the mean and SD of four independent experiments. F: HEK293 cells transfected with the pRL-TK plasmid were treated with various concentrations of PMA as shown for 8 hours. Fold change was calculated by setting the normalized Renilla luciferase RLU for the non-treated sample (0 ng/ml) as 1. The results shown are the mean and SD of three independent experiments.