Sex-related difference in OGD- and OGD/Reox-induced neuronal death in XY and XX CGNs. The cells were exposed to OGD for 2 h or 2 h OGD followed by 6, 12 and 24 h of Reox. A) Morphological evidence of injury in XY compared to that in XX CGNs. A higher magnitude of cell injury was observed in XX CGNs. Degenerated neurons characteristics of apoptosis are shown by yellow arrow head. Scale bar 20 μm. B) Quantification of cell death by LDH release also revealed significantly higher percentage of cell death in female as compared with male CGNs after 6,12 and 24 h Reox following 2 h of OGD. Values are mean ± SD (n = 4), *p < 0.05, **p < 0.01, XX vs. XY, ANOVA followed by Bonferroni/Dunn post hoc test. C) Fluorometric TUNEL immunostaining (green) of CGNs exposed to OGD (2 h)/Reox (6,12 and 24 h) showed significantly enhanced TUNEL (+) cells in XX CGNs during the Reox periods as compared to that observed in XY CGNs (*p < 0.05). Representative images are shown. Scale bar 20 μm.