2.6. Oxygen-glucose deprivation (OGD)

Primary cortical neurons were subjected to OGD insult as reported previously (Frantseva et al., 1999; Newcomb-Fernandez et al., 2001; Yin et al., 2002; Zhao et al., 2009a). Briefly, the neuron culture media were replaced with pre-warmed glucose-deprived Neurobasal-A medium (Invitrogen) pre-equilibrated with 1% oxygen, and then primary neurons were cultured in an incubator filled with 1% oxygen (5% CO₂/94% N₂) for 0.5, 1 or 1.5 h at 37 °C. At the end of hypoxia, cells were transferred to normal culture medium, returned to normal culture condition and incubated for 24 h (reoxygenation).