Figure 3 | Scientific Reports

Figure 3

From: Optimization of BRET saturation assays for robust and sensitive cytosolic protein–protein interaction studies

Figure 3

Monitoring the drug-dependent FRB-FKBP interaction with variable donor BRET saturation assays. (A) Simplified dynamics of the interaction of FKBP with the FRB domain of mTOR mediated by the rapamycin. (B) HEK-293T cells were incubated in presence of rapamycin for 1 h between fluorescence and bioluminescence acquisition. (C) Variable donor BRET saturation assay of FRB-Nluc/YFP-FKBP in the presence or absence of rapamycin. A 3 σ-threshold (0.07) was defined based on SOD-FRB interactions considered a negative control. (D) Variable donor BRET saturation assay of FRB-Nluc/YFP-FKBP in cells treated with various concentrations of rapamycin for 1 h. (E) BRETmax and BRET50 values were plotted according to the different concentrations of rapamycin.

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