With the increasing appreciation of the role of Notch in development and disease, measuring its cleavage and signaling activity in cellular systems has become important. Here we describe a cell-based method to analyze the cleavage of Notch at the S3 site by γ-secretase. HEK cells are transfected with an N-terminal truncated and myc-labeled mNotchΔE construct which can be easily and quantitatively detected by western blotting.
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This work was supported by VIB and a Methusalem grant from KU Leuven and the Flemish Government. We would like to thank Tine Vanhoutvin for useful technical discussions and Veerle Vulsteke for drafting Fig. 1.
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