Fig. 2.

Mutation of E-box 3 significantly decreases basal activity of a mouse GnRHR-Luciferase reporter gene in αT3-1 and LβT2 cells. (A) E-box sequences from the proximal 1000 bp of each gene are shown in comparison to the well-characterized POMC E-box and the Group A consensus (GrpA). Within each gene, both the sense (S) and antisense (AS) strands are shown in the 5’ to 3’ orientation. They are ordered from the most distal (top) to the most proximal (bottom) within each gene. The defining bases (CAnnTG) are in capital letters. The internal base pair matches for the Group A bHLH consensus (GrpA, shown at the top) are underlined. The flanking bp matches for to the well-defined NeuroD1 binding site from the POMC gene (shown second) are in bold. (B) Mutated sequences of E-boxes 3 and 4 from the GnRH gene are shown with the mutated bases underlined. (C) The GnRHR-Luciferase reporter either wild-type or containing mutations in either E-box 4 or E-box 3 were transiently transfected into αT3-1 cells or LβT2 cells.