Figure 3.

Prominin⁺ cells are located in the cerebellar white matter. (a-d) Detection of prominin mRNA by in situ hybridization. Cryosections of P7 cerebellum were hybridized with digoxigenin (DIG)-labeled sense (control; a,b) or antisense (c,d) probes specific for mouse Prom1 (prominin RNA) and then stained with alkaline phosphatase-conjugated antibodies to DIG and with NBT/BCIP substrate to detect bound probe. Sections were photographed at 10× (a,c)and 20× (b,d) magnification; boxes indicate locations of regions shown in b,d. Arrows indicate prominin⁺ cells (dark brown spots) in the white matter (WM). (e,f) Detection of prominin protein by immunostaining. cerebellar sections were stained with rat antibodies to prominin-1 and TRITC-conjugated secondary antibodies. Low-power (10)image of cerebellum from a Math1-GFP mouse shows GFP fluorescence× (green) in the outer EGL and prominin⁺ cells (red) in the white matter (e). The section is counterstained with DAPI (blue) to highlight cerebellar structure. High-power (20×) image of cerebellum from wild-type mouse shows prominin⁺ cells (red) in the white matter (f). No counterstain is shown. Red, green and blue images were photographed separately and merged with Openlab software. EGL, external germinal layer; WM, white matter.