Figure 7

Mesencephalic astrocyte-derived neurotrophic factor expression in cultured primary microglia. Glial cells were cultured as described in Materials and methods. The serum concentration in DMEM was lowered from 10% to 5% during treatment. Immunofluorescent staining was performed in vehicle controls (A to D) and 24 hours after treatment with 1 μg/ml tunicamycin (E to L). Microglia were identified with anti-CD68 antibody (green, A, E, and I). Mesencephalic astrocyte-derived neurotrophic factor (MANF) expression was detected with monoclonal anti-MANF antibody (red, B, F, and J). Nuclei were stained by 4^′,6-diamidino-2-phenylindole (blue, C, G, and K). Scale bar = 50 μm.