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. 2017 Apr 3;7:525. doi: 10.1038/s41598-017-00614-w

Figure 1.

Figure 1

High-throughput luciferase screening to identify parkin inducing compounds. (A) Schematic illustration of the high-throughput screening method. HEK-293T parkin reporter cell line (parkin-Luc-HEK-293T) was selected by stable transfection of the luciferase construct containing three repeats of parkin promoter’s CREB/ATF4 binding motifs. In a 96-well plate, 1172 FDA-approved drugs were used to treat parkin-Luc-HEK-293T cells. Parkin promoter activity was measured by luciferase assay. DMSO was used as a negative control. CCCP treatment was used as a positive control. (B) Parkin promoter activities in parkin-Luc-HEK-293T cell line treated with DMSO or 10 uM CCCP as positive control (n = 4) were measured by luciferase assay. (C) Scatter plot showing relative increase of parkin promoter activity in parkin-Luc-HEK-293T cell line treated with each compound compared to DMSO negative control based on luciferase assay. Top 20 compounds were highlighted in red color. (D) Relative increase of parkin promoter activity induced by the top 20 compounds based on the initial HTS screening determined by luciferase assay (n = 9, see also Table 1). Quantified data are expressed as mean ± s.e.m. *P < 0.05, **P < 0.01, and ***P < 0.001, nonparametric Kruskal-Wallis ANOVA test (B) and ANOVA test followed by Tukey post hoc analysis (D).