Figure 2.

Foxl2 over-expression causes activation of the GnRHR promoter. The GnRHR promoter-firefly luciferase vector (-600 Luc) was co-transfected with either pFoxl2 or pcDNA 3.1. All transfections included the control vector phRLCMV that expresses renilla luciferase to correct for differences in transfection efficiency between samples. Firefly luciferase values were divided by renilla luciferase values to normalize for transfection efficiency. As an additional control, the promoter-less luciferase vector (pGL3 basic) was transfected with either the empty vector pCDNA3 or Foxl2 expression vector (pFoxl2) in order to show that Foxl2 did not affect the luciferase control vector (Data not shown). Data from four independent transfection experiments was combined to generate the graph in Figure 1. Each of the four experiments was performed in triplicate for a total of 12 data points represented in each column. Each of the experiments used different KK1 cell cultures and DNA preparations. Statistical analysis using GraphPad Prism software (paired T-test; p = 0.0067**) allowed us to determine that Foxl2 over-expression caused a 5.8 fold increase in promoter activity.