Biomedical Engineering Reference
In-Depth Information
Fig. 1
Nuclear export and the identifi cation of nuclear export inhibitors. (
a
) The assay is based on a reporter
protein that contains a fl uorescent tag (GFP) and a nuclear export sequence (NES). The export receptor CRM1
recognizes and binds to the NES of the reporter protein and escorts it through the nuclear pore complex. The
export complex is formed by binding RanGTP. Once in the cytoplasm, the export complex dissociates when the
GTP of Ran is hydrolyzed. (
b
) The assay has been adapted to a multiwell format. Treatment of the stable
reporter cell line with inhibitors of the nuclear export that block the interaction between CRM1 and NES-
containing cargo proteins leads to an accumulation of fl uorescent signal in the cell nucleus
2.2 Software
AttoVision software (BD Biosciences, San Jose, CA).
BD Image Data Explorer (Genedata AG, Switzerland).
Genedata Screener software (Genedata AG, Switzerland).
