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Figure 1


Fig. 1. Strategy of Cre-mediated lineage tracing of Notch1 activity. (A) Schematic depicting the mouse Notch1 protein. Indicated are the extracellular domain containing a signal peptide (SP) and 36 EGF-like repeats, the three Lin-Notch repeats (LNR), the transmembrane domain (TM) and the Ram domain, seven ankyrin repeats and the PEST and transcriptional activation domain (Tad-PEST). S2 and S3 indicate Adam metalloprotease-dependent cleavage and the {gamma}-secretase dependent cleavage at Val1744, respectively. Using gene-targeting, Cre recombinase was inserted immediately downstream of Val1744 at Arg1752. Interaction of N1::cre receptors in vivo with Notch DSL ligands results in S2 and S3 proteolytic cleavages and release of Cre recombinase from the plasma membrane. (B) Cre recombinase can irreversibly activate the ubiquitously expressed R26R reporter and permanently mark cells with lacZ expression in vivo. (C) If N1::cre is activated in a stem cell, all surviving descendents appear blue; when Notch1 is activated in progenitors or differentiated cells, a mixture of blue and white cells will appear in any given tissue. (D,E) Sagittal view of whole-mount X-Gal staining of E14.5 N1::cre;R26R embryos showing identical patterns of widespread labeling of several tissues. Black arrows indicate strong thymic staining and white arrows indicate dorsal aorta (D) and umbilical artery (E). Magnification: 10x.





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