Figure 6.

Evaluation of DBD mutant ERα activities in vivo and in vitro. A, Uterine cross sections from ovariectomized WT, KIKO, EAAE, or ERα-null mice that were treated for 24 hours with E₂. Sections were evaluated for the proliferative marker, Ki67 (brown). Scale bar corresponds to 0.1 mm. B, WT, KIKO, and EAAE ERα mediate AP1-luc responses similarly. WT, KIKO, and EAAE ERα with AP1-luc. Fold changes relative to vehicle (V) treatment are indicated above each bar. E₂, 10 nM; ICI 182,780 (ICI), 100 nM. Statistical analysis included 2-way ANOVA, multiple comparisons of means vs results for vehicle treatment, and Bonferroni multiple test correction. ***, P < .001; ****, P < .0001. C, Microarray profile. A hierarchical cluster of normalized ratios (E₂ 2h/V) of WT and EAAE uterine RNA is shown. Uterine RNA from WT or EAAE ovariectomized mice collected 2 hours after vehicle (V) or E₂ injection (n = 3 each condition) was compared by microarray. The cluster was created by combining replicates and building ratios (E₂ treated/vehicle treated) and filtering for probes that met the following criteria: absolute fold change of > 2 in at least 1 genotype, probe intensity of >100 in at least 1 condition, and false discovery rate < 0.05. This resulted in 3162 probes. Red indicates induced transcripts; green signifies repression.