Figure 1.

The experimental timeline of oxygen and glucose deprivation/reoxygenation (OGD/R) exposure and rMS stimulation. On day 0, the equal amount of Neuro-2a (N2a) cells were seeded in growth medium, which contains 10% fetal bovine serum (FBS) and 1% Penicillin–Streptomycin solution. At 70–80% confluency, the growth medium of N2a cells was changed into differentiation medium, which contains 20 µM of retinoic acid (RA) in Dulbecco’s Modified Eagle Medium (DMEM) for 4 days. After washing three times with PBS, the differentiation medium of N2a cells was changed into deoxygenated, glucose-free balanced salt solution (Gibco) and placed into a hypoxic chamber (O₂ tension 1%) for 3 h. After this OGD injury, the cells were placed with medium and received on–off interval of 3 s stimulation for 10 min. After the stimulation, the cells were incubated in a humidified atmosphere of 5% CO₂ at 37°C. After 24 h, OGD/R+rMS cells were harvested for cell proliferation analysis, qRT-PCR, Western blot, ICC, and TUNEL assay. RA, retinoic acid; OGD/R, oxygen and glucose deprivation/reperfusion; rMS, repetitive magnetic stimulation; qRT-PCR, quantitive real-time reverse transcription polymerase chain reaction; ICC, immunocytochemistry; TUNEL, terminal dUTP nick end-labeling.