Cellular and Biophysical Pipeline for the Screening of Peroxisome Proliferator-Activated Receptor Beta/Delta Agonists: Avoiding False Positives

Figure 3

Development of a cytotoxic parameter. Comparison between the Renilla reporter expression with the MTT cytotoxicity signal in different DMSO concentrations (0.1%, 1%, 3%, 5%, 10%, and 50%) in order to validate the Renilla reporter expression as an indirect cytotoxicity parameter. One micromolar GW0742 diluted in DMSO, with a final concentration of 1% DMSO, was used as the positive control in the screening. Statistical analyses proved that our indirect cytotoxic parameter (Renilla reporter expression) can be used to identify cytotoxic compounds as well as the MTT cytotoxicity assay. Through analyzing the 2-way ANOVA statistics, there is no statistical difference (ns) between the two types of experiments, and the effect of the DMSO concentration was considered extremely significant according to GraphPad Prism. Followed by Sidak’s multiple comparison test, the toxicity increases in concentrations above 3% of DMSO with statistical significance. Data are the mean ± SD ( independent replicates).