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. 2020 Jul 31;2(4):e190053. doi: 10.1148/rycan.2020190053

Figure 6:

Reversible notch signaling reporter. A, In the endogenous Notch pathway, ligand binding leads to ectodomain shedding due to cleavage at the S2 site by ADAM, followed by intramembrane proteolysis at the S3 site by γ-secretase. The released Notch intracellular domain (NICD) translocates to the nucleus and interacts with the DNA-binding protein CSL to recruit MAML proteins and other coactivators to activate gene expression. In the split luciferase construct, each half of the luciferase is kept separated and dark until the pathway is fully engaged in the nucleus. B, The reporter recapitulates the dose response curve of the Notch pathway to DAPT both linearly and over a large dynamic range. This inhibition could be specifically sensitized with siRNA targeting NCSTN. (Reprinted, with permission, from reference 49.) CSL = CBF1/RBPjκ/Su(H)/Lag1, DMSO = dimethyl sulfoxide, LCI = luciferase complementation imaging, MAML = mastermind like.

Reversible notch signaling reporter. A, In the endogenous Notch pathway, ligand binding leads to ectodomain shedding due to cleavage at the S2 site by ADAM, followed by intramembrane proteolysis at the S3 site by γ-secretase. The released Notch intracellular domain (NICD) translocates to the nucleus and interacts with the DNA-binding protein CSL to recruit MAML proteins and other coactivators to activate gene expression. In the split luciferase construct, each half of the luciferase is kept separated and dark until the pathway is fully engaged in the nucleus. B, The reporter recapitulates the dose response curve of the Notch pathway to DAPT both linearly and over a large dynamic range. This inhibition could be specifically sensitized with siRNA targeting NCSTN. (Reprinted, with permission, from reference 49.) CSL = CBF1/RBPjκ/Su(H)/Lag1, DMSO = dimethyl sulfoxide, LCI = luciferase complementation imaging, MAML = mastermind like.