Fig. 4

ERα mRNA expression increased in female hippocampal neurons following OGD/REOX and was blocked by ANA-12 treatment or T pretreatment. At the end of REOX, cells were either stained for ERα or harvested for mRNA extraction and probed for mRNAexpression of ERα. A. Representative images of female hippocampal neurons stained for ERα (green) and counterstained with MAP-2 (red) and DAPI (blue) after 4 h OGD and 24 h REOX. Arrow: ERα nuclear staining. Arrowhead: ERα neurite staining. B. Summary figure of relative ERα mRNAexpression in cultured hippocampal neurons under normoxic conditions and after exposure to 4 h OGD and 3 h REOX. Cells were treated with either vehicle control, 3 µM 7,8-DHF, 100 µM ANA-12 or 3 µM 7,8-DHF + 100 µM ANA-12 during REOX. Values are expressed relative to male normoxia. Values are mean ± SEM, n = 3–12. * p = 0.009 vs. female 4 h OGD and 3 h REOX; # p = 0.02 vs. female 4 h OGD and 3 h REOX + 3µM 7,8-DHF. Significance was determined by multi-factorial analysis of variance. C. Relative ERα mRNA expression in cultured hippocampal neurons under normoxic conditions and after 4 h OGD and 3 h REOX. Cells were pretreated with either vehicle control (VC) or 10 nM T (T). Values are expressed relative to male normoxia. Values are mean ± SEM, n = 3–8. * p = 0.0001 vs. VC treated female 4 h OGD and 3 h REOX. Significance was determined by multi-factorial analysis of variance