Figure 1

Mesencephalic astrocyte-derived neurotrophic factor expression in the astrocytes of brain tissue. Brain samples were collected from the normal (A to C) and ischemic rat brain tissue (D to L). (D to F) and (J to L) Two-hour ischemia followed by 24-hour reperfusion; (G to I) 4-hour ischemia followed by 24-hour reperfusion. (A to I) Mesencephalic astrocyte-derived neurotrophic factor (MANF) expression in astrocytes was detected by double immunofluorescent staining for glial fibrillary acidic protein (GFAP) (red, A, D, and G) and MANF (green, B, E, and H) with anti-GFAP and anti-MANF, respectively. (J to L) MANF expression in neurons was detected with anti-MANF (red, K). The neurons were identified by anti-NeuN (green, J). Ischemia-induced MANF expression was found predominantly in neurons (K). A small number of astrocytes expressing MANF are indicated by arrows. Scale bar = 50 μm. (M to P) Specificity of the antibody against MANF in brain tissues was detected by immunohistochemistry and immunofluorescent staining. Brain sections were incubated with primary antibody (anti-MANF) in (N) and (P). Negative controls were performed by substituting the primary antibody with PBS (M and O). Scale bar = 50 μm.