Figure 7.

Hypoxia–reoxygenation induces translation of PrP^C and ERK1/2 in primary cortical cultures. A, primary cortical cultures were placed in a hypoxic chamber (95% N₂ and 5% CO₂, 37°C) for 12 h and returned to a normoxic incubator (95% air and 5% CO₂, 37°C) for different lengths of time (1, 3, 10, and 24 h), after which, total protein was extracted for Western blot analysis of PrP^C. B, Densitometric analysis of the Western blot, the ratio of PrP^C/actin was taken as 1 for nonhypoxic cells, which represents the control (C). The mean ± SEM is shown. *p < 0.05 versus control. The time-dependent changes in PrP^C expression are depicted. C, Primary cortical cultures were treated as above, and total protein was then extracted for Western blot analysis of ERK1/2, p38, and JNK expression. D, Densitometric analysis of the Western blot, the ratio of ERK1/2, p38, and JNK/actin were taken as 1 for nonhypoxic cells, which represents the control. E, Primary cortical cultures were treated as above and total protein was then extracted for Western blot analysis of PrP^C expression with or without addition of inhibitor of PD98059. F, Densitometric analysis of the Western blot, the ratio of PrP^C/Actin were taken as 1 for nonhypoxic cells, which represents the control. G, The results show that N18 cells infected with rAd-PGK-PrP^C-Flag subjected to hypoxia for 12 h exhibited significantly reduced caspase-3 activity compared with that of rAd-PGK-eGFP. H, The rAd-PGK-PrP^C-Flag-infected N18 cells showed much reduced LDH activity after 24 h of hypoxic treatment compared with that of rAd-PGK-eGFP. The mean ± SEM is shown. ^*p < 0.05 versus control. C, Control.