Fig. 3

7,8-DHF-mediated neuroprotection was dependent on ERα in cultured hippocampal neurons. ERα^+/+ and ERα^−/− hippocampal neurons were grown on coverslips and 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 cells respectively. A. Representative images of male and female ERα^+/+ hippocampal neurons under normoxic conditions or after 4 h OGD followed by 24 h REOX with and without 3 µM 7,8-DHF treatment. Arrow = Hoechst 33349 nuclear staining (all cells); Arrowhead = co-localized Hoechst 33349 and PI (dead cells) B. Representative images of male and female ERα^−/− hippocampal under normoxic conditions or after 4 h OGD followed by 24 h REOX with and without 3 µM 7,8-DHF treatment. C. Summary figure of % cell survival in ERα^+/+ hippocampal neurons subjected to 4 h OGD and 24 h REOX. OGD resulted in a decrease in cell survival in male and female ERα^+/+ hippocampal neurons. Treatment with 3 µM 7,8-DHF rescued female ERα^+/+ hippocampal neurons, but not male neurons. Data are mean ± SEM, n = 5–7. D. Summary figure of % cell survival in ERα^−/− hippocampal neurons subjected to 4 h OGD and 24 h REOX. OGD resulted in a decrease in cell survival in male and female ERα^−/− hippocampal neurons. However, treatment with 3 µM 7,8-DHF failed to rescue either male or female ERα^−/− hippocampal neurons. Data are mean ± SEM, n = 3–6. * p = 0.004 vs. female ERα^+/+ OGD/REOX + 7,8-DHF. Significance was determined by multi-factorial analysis of variance