Fig. 3. BAD-LAMP is specifically expressed in neurons of the cortical layers II, III and V. (A) In situ hybridization for Bad-lamp (top panel), Cux2 (middle) and ER81 (bottom) on adult mouse brain coronal hemisections. (B) High magnification views of in situ hybridization on wild-type cortex shown in A. Bad-lamp is expressed in layers II, II and V, but excluded from layers IV and VI. In the Scrambler cortex, the entire region appears disorganized. However, the typical inversion of cortical layers is reflected by the altered BAD-LAMP staining, demonstrating that projection neurons express the protein. (C) Combined in situ hybridization for Bad-lamp (in blue) with immunohistochemistry for the specific neuronal marker NeuN (in brown). The left panel is a higher magnification of the boxed area. Bad-lamp is co-expressed with this pan-neuronal marker in many, although not all, neurons. (D) Immunohistochemistry of BAD-LAMP in the indicated cortex layers. (E) Immunofluorescence staining on adult cortex using anti-MAP2 (green) and anti-BAD-LAMP (red) antibodies; the merged image is on the left. Whereas MAP2 is present along the entire dendrites, BAD-LAMP accumulates in defined domains (arrows). Bars, 500 µm in A; 200 µm in B; 100 µm (left panel) and 10 µm (right panel) in C; 10 µm in D.