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Blood, Vol. 113, Issue 18, 4381-4390, April 30, 2009
Structure of the Notch1-negative regulatory region: implications for normal activation and pathogenic signaling in T-ALL
Blood Gordon et al.
113: 4381
Supplemental materials for: Gordon et al
Files in this Data Supplement:
- Figure S1. Effect of the D1561A mutation in LNR-C on reporter gene activity (JPG, 40.2 KB)
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The calcium coordinating residue D1561 was mutated to alanine, and the effect of this mutation in the context of a Notch1 polypeptide lacking the EGF-like ligand binding repeats (ΔEGF) was examined in a reporter gene assay. The reporter activities of ΔEGF alone, ΔEGFΔLNR, and full-length Notch1 were also assessed for comparison. The receptors were transiently transfected into U2OS cells together with a plasmid encoding a luciferase reporter gene under control of four iterated CSL binding sites, and an internal control plasmid expressing Renilla luciferase. Firefly luciferase activities were normalized to the internal Renilla control and expressed relative to the activity of transfection of the pcDNA3 empty vector, which was assigned a relative value of 1.
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