Effects of Nrdp1 on OGD-induced apoptosis in primary rat cerebral cortical neurons. (A) The infection efficiency of neurons with Ad-control, Ad-Nrdp1, Ad-si-control and Ad-si-Nrdp1 was visualized for green fluorescent protein (GFP) 24 h later using fluorescence microscopy (magnification, ×400). (B) Western blot analysis showed that incubation neurons with Ad-si-control and Ad-si-Nrdp1 for 48 h significantly (~90%) reduced Nrdp1 protein levels. Upper panel: representative immunoblots of Nrdp1 and the loading control β-actin; bottom panel: quantitative data of protein band intensity after normalization to β-actin. *P < 0.05 vs. Ad-si-control, ANOVA; n = 4. (C) Western blot analysis showed that incubation neurons with Ad-control and Ad-Nrdp1 for 48 h significantly increased Nrdp1 protein levels. Upper panel: representative immunoblots of Nrdp1 and the loading control β-actin; bottom panel: quantitative data of protein band intensity after normalization to β-actin. *P < 0.05 vs. Ad-control, ANOVA; n = 4. (D) Neurons were infected by with Ad-control, Ad-Nrdp1 or Ad-si-Nrdp1 and then treated with OGD for 6 h. Cell death rate was assessed by lactate dehydrogenase (LDH) release. *P < 0.05 vs. Ad-control. #P < 0.05 vs. Ad-control + OGD, ANOVA; n = 4. (E) Apoptosis was detected using TUNEL assay. Quantitative analysis of TUNEL-positive cells from three independent experiments. *P < 0.05 vs. Ad-control. #P < 0.05 vs. Ad-control + OGD, ANOVA; n = 4. (F) Neurons were infected and treated with OGD as in (D). Western blots analysis of expression of cleaved PARP protein (upper panel). Quantitative analysis of cleaved PARP was shown in the bottom panel. *P < 0.05 vs. Ad-control. #P < 0.05 vs. Ad-control + OGD, ANOVA; n = 4. (G) Western blots analysis of expression of Bax and Bcl-2 proteins (upper panel). Quantitative analysis of the ratio of Bax/Bcl-2 was shown in the bottom panel. *P < 0.05 vs. Ad-control. #P < 0.05 vs. Ad-control + OGD, ANOVA; n = 4.