Figure 1.
Neuroprotective effect of AIF knockdown against oxygen–glucose deprivation injury in primary hippocampal/cortical neurons. A, Western blots show that AIF expression was decreased time dependently in neurons infected with AAV expressing the AIF targeting sequence (AAV–AIFt) (Table 1). The expression levels of COX IV, VDAC, and HSP60 were unchanged. B, Semiquantitative data of A, based on four independent experiments. *p < 0.05, ***p < 0.001 versus empty AAV-infected cells. C, Confocal images showing AIF expression (red) in neurons coinfected with AAV–AIFs (scramble control sequence) and AAV–GFP (a–c) or AAV–AIFt and AAV–GFP (d–f). The arrows in e and f point to infected neurons showing decreased AIF expression. D, Hoechst 33258 (top) and dual TUNEL/Cyto11 green fluorescent nuclear staining (bottom) in normal unstressed neurons (a, d) and neurons at 24 h after 60 min of OGD (b, c, e, f). After OGD, many neurons showed condensed nuclei (b) and TUNEL-positive nuclei (yellow–red, e); condensed nuclei and TUNEL-positive nuclei were present less frequently in cells infected with AAV–AIFt (c, f). E, Percentages of condensed nuclei were quantified at 24 h after OGD of different duration in neurons either nontransfected or infected with AAV–AIFs or AAV–AIFt. F, Cell viability was measured based on Alamar blue fluorescence at 24 h after OGD of different duration. G, Cell viability was measured at 48 and 72 h after OGD (60 min). *p < 0.05, **p < 0.01 versus nontransfected neurons. Data are mean ± SE; n = 12 per experimental condition from three independent transfection experiments.