Fig. 1

Characterization of the mouse primary cortical neuron and BV2 microglia co-cultures and effects of neuroinflammation. a Immunofluorescence staining showed healthy MAP2-positive cortical DIV7 neurons in BV2 microglia co-cultures. Upper panel shows MAP2-positive (green) neurons and GFAP-positive (red) astrocytes. Lower panel shows CD11b-positive (red) BV2 microglia cells (white arrow) in MAP2-positive cortical neuron (green) and BV2 cell co-culture. Nuclei are stained with DAPI. Magnification × 20, scale bar 50 μm. The quantification of the data shows that the ratio of astrocytes to neurons (white column) is significantly increased in the co-cultures compared to neurons only cultures. This might reflect either a true increase in the amount of astrocytes or decreased amount of neurons due to addition of microglial cells. Black column indicates the ratio of BV2 microglial cells to neurons at DIV7. b LPS and IFN-γ treatment was used to induce neuroinflammation in mouse primary cortical neuron and BV2 microglia co-cultures. Neuronal viability decreased by 60% after induction of neuroinflammation. Pretreatment with iNOS inhibitor 1400 W (20 μM) prevented neuronal loss and production of NO. Anti-inflammatory cytokine IL10 decreased TNFα levels, but had no effect on viability or NO levels. *p < 0.05, **p < 0.01, ***p < 0.001, mean ± SEM, n = 23 (a), and n = 5–6 (b)