Figure 7.

Effects of siRNA targeting Nix or VEGF on cellular apoptosis after ischemic challenge. A. siRNA targeting Nix, VEGF or luciferase was transfected into primary cortical neurons 30 h before OGD. TUNEL staining was performed 96 h after OGD challenge. TUNEL‐positive cell counting analysis was determined under each condition. Quantification of TUNEL‐positive cells was presented as mean (SD) (n = 4). Scale bars, 50 µm. B. Nix or VEGF in OGD‐treated or untreated primary neurons was knocked down by using the appropriate siRNA. The viability of the cultured cortical neurons was assessed by performing the MTS assay. The Nix and VEGF expression levels were used as an internal control for the MTS assay. Experiments were carried out independently in triplicate. The values that were normalized to the protein content are provided as the means (SD) (n = 3) and have been expressed as percentage viability relative to the untreated cells. Statistical analysis was carried out using the one‐way ANOVA with appropriate post hoc tests; *P < 0.05 vs. the naive‐control group. †P < 0.05 vs. OGD plus luciferase siRNA group. Abbreviations: siRNA = small interfering RNA; Nix = Nip‐like protein X; VEGF = vascular endothelial growth factor; OGD = oxygen–glucose deprivation; TUNEL = terminal deoxyuridine‐triphosphate nick‐end labeling; MTS = methanethiosulfonate; SD = standard deviation.