Fig. 6. Analysis of Notch 1 ICD and SMAD1-responsive elements in the Hey1 promoter. (A) Schematic picture of the Hey1 reporter construct with CSL-binding sites and the GC-rich domain (Hey1-luciferase), and the two mutants containing only the GC-rich domain (GC-luciferase) and comprising the Hey1 promoter lacking the GC-rich domain (Hey1-GC). (B) Transfection of Notch 1 ICD (67 ng), SMAD1 (67 ng) and/or R218H CSL (67 ng) in COS-7 cells, in the absence (white bars) or presence (black bars) of exogenous BMP4 (50 ng/ml), measured by activation of the GC-luciferase reporter construct. (C) Transfection of Notch 1 ICD (100 ng) and SMAD1 (100 ng) in COS-7 cells, in the absence (left) or presence (right) of exogenous (50 ng/ml) BMP4, measured by activation of Hey1-luciferase (white bars) or Hey1-GC (black bars) reporter constructs.

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