Figure 2.

Cell type and tissue-specific expression of NPC1-YFP. A, Immunofluorescent confocal analysis of NPC1-YFP (GFP) localization within astrocytes (S100) or PNs (D28K) in the cerebellum. NPC1-YFP localizes to astrocytes in a G; N mouse and within PN in a P; N mouse (arrows). NPC1-YFP did not localize to other cells. Nuclei were labeled with Hoechst. B, Epifluorescent microscopy showing the distribution of NPC1-YFP relative to PNs. NPC1-YFP colocalizes with PNs in a P; N but not in a G; N mouse cerebellum. C, Immunoblot analysis shows expression of NPC1-YFP throughout the brain in G; N mice. Expression of NPC1-YFP in P; N mice is restricted to the cerebellum. Anti-actin was used as a loading control. D, Immunofluorescent confocal microscopy shows localization of NPC1-YFP (GFP) to MAP2-positive cells in the thalamus of an E; N mouse. E, For contrast, grayscale, inverted images were created using Photoshop. Black pixels are positive for fluorescence, gray is tissue background, and white represents no signal. Immunofluorescent signal from sagittal brain sections showed strong NPC1-YFP accumulation in the striatum (STR) and in the PN layer (arrow) of the cerebellum (CB). Relatively, weak NPC1-YFP immunofluorescence is detectable in the cortex (CTX). The level of NPC1-YFP in the cortex is representative of other CNS regions that are not shown. F, In a C; N mouse, the predominant NPC1-YFP expression is localized to the forebrain areas including hippocampus (HP), but little and sparse NPC1-YFP is found in the thalamus (TH). Also, the majority of cerebellar PNs do not produce NPC1-YFP in this mouse strain. G, NPC1-YFP is detected only in the olfactory bulb (OB) and choroid plexus (CP, arrow) of the brain of R; N mice. Aside from brain, NPC1-YFP is detected in multiple viscera tissue. Sections of liver and intestine are shown.