Figure 1.

Diagram of the mNotch1 gene product and retroviral constructs. The molecule consists of an extracellular domain containing 36 tandem epidermal growth factor (EGF) repeats and 3 lin-12/Notch (LNR) repeats; a single transmembrane domain (TM); and an intracellular domain containing 6 cdc10/SWI6/ankyrin repeats, putative nuclear localization signals (NLS) and a C-terminal OPA/PEST region. The activated mNotch1 construct (MT-mNotch-ICΔOP) consists of the region of the intracellular domain including the cdc10 repeats and 3′ nuclear localization signals. The constructs included 5′ myc epitope tags (MT) to facilitate detection of protein expression. The control construct MT-mNotch1-ICΔOPΔCDC lacks the 5′ end of mNotch through the cdc10 repeats, but otherwise corresponds to MT-mNotch1-ICΔOP. Constructs were cloned into the LXSN retroviral vector that had been modified to contain a ClaI restriction site.