Figure 3

Induction of Noxa and effect of noxa deletion on neuronal injury primary cortical neurons. (a) Western blotting and densitometric analysis comparing the levels of Noxa induction in OGD-treated cortical neurons allowed to recover under normoxic conditions for the times indicated, confirming significant Noxa protein induction. Significant increases in Noxa induction were observed at 0 h and maintained up to 24 h compared with normoxic controls. Data presented as mean±S.E.M. from n=4 independent experiments from n=4 independent cultures. *P<0.05 compared with sham-treated controls. (b and c) Cortical neurons derived from WT and noxa-deficient mice were sham-treated or subjected to either 45 min OGD or a model of excitotoxic NMDA receptor overactivation by exposure to NMDA/glycine (100 μM/10 μM) for 5 min and allowed to recover under normoxic conditions for 24 h. Cell death was assessed after 24 h in each model by Hoechst and PI staining, and three subfields were captured per well, with a minimum of three wells analyzed per condition. Nuclei uniformly stained with Hoechst were counted as viable and condensed; PI-positive nuclei were scored as dead neurons and expressed as a percentage of the total population. Deficiency of noxa in cortical neurons neither reduced (b) OGD- or (c) glutamate-induced neuronal injury in cortical neurons. Data presented as mean±S.E.M., figures representative of n=3 independent experiments from n=3 independent cultures carried out in triplicate with similar results. *P<0.05 compared with sham-treated control; ^#P<0.05 between treatments (ANOVA, post hoc Tukey’s test)