Figure 4.

 The effect of PCDH19 variants on ERα-mediated gene expression. (A) Luciferase activity was assayed in MCF-7 cells transfected with reporter containing three copies of vitellogenin Estrogen Response Element (3× ERE TATA luc) and either control, wild-type (WT) or mutant (MT) Myc-PCDH19 expression vectors. Cells were initially cultured in charcoal-stripped FCS medium for 16 h and then for 6 h in the presence or absence of 10 nM estradiol (E2). Data are expressed as relative luciferase activity ± SD from four or more independent experiments. **P < 0.01 comparing +E2 versus–E2 vector control, **¹P < 0.01 comparing +E2 versus –E2 WT, **²P < 0.01 comparing +E2 and –E2 WT versus vector or **³P < 0.01 comparing –E2 WT versus –E2 MT. **⁴P < 0.01 comparing +E2 WT versus  +E2 MT using Bonferroni adjusted planned comparisons. (B) Levels of PCDH19 and endogenous-NONO protein were determined by western blotting with anti-Myc and anti-NONO antibodies. β-Actin was used as a loading control.