Figure 3. CaMKII inhibition attenuates apoptosis in rat hippocampal slice cultures exposed to oxygen glucose deprivation.

Rat hippocampal slice cultures were exposed to OGD in the presence of the CaMKII inhibitor, KN93 or its inactive analog, KN92 (10 µM, 2 h prior to OGD). Slices were harvested 8 h after OGD and subjected to Western blot analysis to determine effects on cleaved caspase-3 (A). A representative blot is shown (A). OGD increases cleaved caspase-3 levels and this is attenuated by KN93, but not by KN92 (A). Slices were also subjected to TUNEL analysis. Representative images are shown demonstrating TUNEL staining of apoptotic cells (green) co-localized with PI staining of all the nuclei (red) (B). The magnification used was 10×. Quantification of the percentage of apoptotic nuclei to total nuclei was also carried out indicating that KN-93 pretreatment decreased the level of apoptotic nuclei in response to OGD (C). Data are presented as mean ± S.E from 4 independent experiments using 24 pooled slices per experiment. * P<0.05 vs. no OGD, † P<0.05 vs. OGD alone, ‡P<0.05 vs. OGD+KN92.