WT1 (+KTS) decreases basal and GnRH-stimulated LHβ promoter activity.

<p>LβT2 cells were transfected with either <b>A)</b> A luciferase reporter construct driven by the rat LHβ promoter (-617 to +41 bp) including both distal and proximal GnRH responsive promoter regions, or <b>B)</b> A luciferase reporter construct driven by the rat LHβ promoter (-245 to +44 bp), including only the proximal GnRH response region of the promoter. Constructs were cotransfected with or without 0.5, 0.7,1 μg of WT1 (+KTS) plasmid or control plasmid to normalize DNA. At 48 h post-transfection, cells were treated with 50nM GnRH for 6 h and collected in lysis buffer. Luciferase activity was measured, and data expressed as average ± SE for 6 samples; the experiment was performed 3 times each. Statistical significance was determined using ANOVA (confidence interval determined by the Bonferroni multiple comparison test). * p<.05 -GnRH vs +GnRH, # = p<.05 control, -GnRH vs –GnRH+WT1, a = p<.05 control, +GnRH vs +GnRH +WT1.