Figure 8.

NeuN, GFAP, MAP2, and NF200 expression in the developing cortex. A, B, Representative photomicrographs of WT and Manf^-/- cortical sections stained with NeuN (A) or MAP2 (B) at E19, and counterstained with DAPI (blue) at E19. C, The fluorescent intensities divided by the area of CP. There is no difference between WT and Manf^-/- embryos in the NeuN and MAP2 fluorescent intensities/area of CP (n = 3). D, E, Western blotting from E19 cortical lysates stained with antibodies against NeuN, MAP2, GFAP, and NF200 (n = 4, *p < 0.05). F, Representative photomicrographs of anti-NeuN (green) antibody-stained cortical sections of WT and Manf^-/- mice at P7, counterstained with DAPI (blue). G, The density of NeuN-positive neurons in cortical layers at P7. In the Manf^-/- mouse brain there is a significant increase in NeuN-positive neuronal density in the Layers II–IV and V at P7 (n = 3, *p < 0.05, **p < 0.01). H, Representative photomicrographs of WT and Manf^-/- cortical sections stained with GFAP. I, Representative photomicrographs of WT and Manf^-/- cortical sections stained with MAP2. J, Shorter MAP2-stained dendrites were found in Manf^-/- cortical neurons at Layer IV/V of cortex compared to those in WT (n = 3, *p < 0.05). K, Layer V/VI of cortex from Manf^-/- and WT littermates stained with neurofilament antibody (NF200). Cell nuclei were counterstained with DAPI. L, At P7, there is no difference in immunofluorescence intensities of MAP2 and GFAP-stained cortical sections between groups. However, there was a significant decrease in axon-specific neurofilament immunoreactivity in Manf^-/- cortex (n = 3, *p < 0.05). M, N, Western blotting from P7 cortical lysates probed with glial and neuron-specific antibodies. Decreased levels of NF200 was detected in the cortical extracts from Manf^-/- mice (n = 4, *p < 0.05); scale bars: 50 µm (A, B, F, H) and 25 µm (I, K).