Fig. 5.

Nur77 transcription stimulating activity induced by GnRH requires PKC but not ERK signaling. An expression vector encoding a Gal4 DNA binding domain fused in frame with full length Nur77 was cotransfected into αT3-1 cells along with a luciferase reporter containing five Gal4 DNA binding sites upstream of luciferase. Transfected cells were pretreated with dimethylsulfoxide (vehicle), GFX109203X (GFX; 1 μm) or PD98059 (50 μm) for 30 min followed by application of either saline or GnRHa for 6 h. Luciferase activity was then determined in cell lysates. Bars represent mean ± sem normalized luciferase activity carried out in triplicate within a single experiment. Each experiment was repeated three times with similar results. Bars with different letter designations represent mean values that are statistically significantly different at P < 0.05.