Figure 2. Inhibition of aNF-κB promotes survival of retinal ganglion cells in vitro after OGD.

(A–C) Quantification of active caspase-3 positive RGCs in co-culture with astrocytes 24 hours after OGD challenge (OGD) vs. normoxic controls (Control) performed by confocal microscopy. The total quantities of NeuN or beta-III tubulin/Hoechst positive RGCs observed in 5 randomly selected fields of each experimental plate are compared. Values are means ± SEM, n=5; *p<0.05. (A) Activated caspase-3 positive RGCs in co-culture with WT and TG astrocytes; (B) Activated caspase-3 positive WT RGC in co-cultures with astrocytes treated with the NADPH oxidase inhibitors DPI (1µM) or apocynin (100µM); (C) Activated caspase-3 positive RGCs in co-cultures of WT astrocytes with WT or p47^null RGCs (D) Cell death rates in WT and p47^null RGCs challenged by astrocyte-conditioned media (ACM) from primary astrocytes subjected to OGD challenge (OGD ACM) or from normoxic control astrocytes (Control ACM). The percentages of dead (PI/Hoechst positive) and apoptotic (Annexin V/Hoechst positive) RGCs in ACM-treated cultures were calculated relative to total quantities of Hoechst positive cell in 5 randomly selected fields. Values are means ± SEM, n=5; *p<0.05.