Figure 1 | Scientific Reports

Figure 1

From: Activators of alpha synuclein expression identified by reporter cell line-based high throughput drug screen

Figure 1

SNCA-GFP-LUC reporter cell line-based high throughput screening assay of 1649 FDA approved drugs identifies modulators of α-syn mRNA and protein levels. (A) Schematic overview of CRISPR/Cas9 generated, genomic SNCA-GFP-LUC fusion under control of the endogenous SNCA promotor in the SH-SY5Y cell line. Translation of its mRNA results in both, α-syn-GFP fusion protein and a functional LUC protein, separated by a ribosome skipping event due to the T2A sequence. (B) Agarose gel-electrophoresis of PCR from cDNA (top) and Western blot (bottom) of the reporter cell line confirming in-frame SNCA-GFP-LUC fusion at mRNA- and α-syn-GFP at protein level, respectively. (C) Workflow of the screening and validation procedure of α-syn modulating drugs. Several exposure images of full-size WB in (B) (lower panel) see Supplementary Fig. S2.

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