Fig. 5

T pretreatment in ERα^+/+ hippocampal neurons inhibited 7,8-DHF mediated neuroprotection following OGD/REOX ERα^+/+ and ERα^−/− hippocampal neurons pretreated with either VC or 10 nM T were subjected to 4 h OGD followed by VC or 3 µM 7,8-DHF during 24 h REOX. Cells were stained with Hoechst 33349 and PI to detect total and dead respectively. (A) Representative images of male and female T or vehicle control pretreated ERα^+/+ hippocampal neurons under normoxic conditions or after 4 h OGD followed by VC or 3 µM 7,8-DHF during 24 h REOX. Arrow = Hoechst 33349 nuclear staining (all cells); Arrowhead = co-localized Hoechst 33349 and PI (dead cells) (B) Summary figure of % cell survival in vehicle control (VC) and 10 nM T pretreated ERα^+/+ hippocampal neurons subjected to 4 h OGD and VC or 7,8 µM-DHF during 24 h REOX. OGD resulted in a decrease in cell survival in male and female ERα^+/+ hippocampal neurons. Following T pretreatment, 7,8-DHF failed to rescue either female ERα^+/+ hippocampal neurons. Data are mean ± SEM, n = 5. Significance was determined by multi-factorial analysis of variance