NICD generation involves a series of proteolytic cleavages. Initially, the Notch receptor is synthesized as a single polypeptide and then cleaved in the Golgi apparatus to form a heterodimer. Upon ligand binding, a second cleavage occurs at the extracellular domain by an ADAM-family metalloprotease. The final cleavage is facilitated by the γ-secretase complex, releasing the NICD into the cytoplasm.