Figure 6. T₃ effects on TSHα promoter activity and histone H3 modifications in α-23 cells overexpressed with wild-type TRβ and dominant negative mutant Mf-1.

A) TSHα promoter activity in α-23 cells transformed with adenovirus green fluorescent protein (Ad-GFP), adenovirus TRβ (Ad-TRβ) and adenovirus Mf-1. α-23 cells were plated in 12 well plates and cultured in charcoal-stripped 10% FBS-DMEM for three days. Cells were transformed with adenovirus TRβ or Mf-1 on the fourth day. On the fifth day, cells were treated for 1 hr -/+ 0.1 µM T₃ before harvest and measurement of luciferase mRNA expression by quantitative RT-PCR. B) Histone H3 modifications on TSHα promoter in α-23 cells transformed with adenovirus expressing GRP, TRβ, or Mf-1. α-23 cells transformed with adenovirus GFP, TRβ, or Mf-1 were treated for 1 hour with −/+ 0.1 µM T₃ before harvest and ChIP assay. Shown are quantitative RT-PCR analyses of PCR products from using antibodies directed against specific H3 modifications as indicated in the Figure (n = 3 experiments). Statistical analyses performed as in Figure 1. **, p<0.05; *, p<0.01 from untreated and treated controls, respectively using ANOVA analyses.