Figure 7. CB ₁ levels are reduced in NPC mouse model and patient brains, and FAAHi prevents aberrant phenotypes in NPC cells and mouse model.

• A
  Western blot against CB₁ and GAPDH (used as loading control) and graph showing mean ± SEM CB₁ protein levels in cerebellar extracts from WT and Npc1^nmf164 mice (n = 6).
• B
  Immunofluorescence images against CB₁ and calbindin in the cerebellum of WT and Npc1^nmf164 mice. DAPI stains cell nuclei. Graphs show mean ± SEM intensity associated with CB₁ in the Purkinje cells (indicated by arrows) expressed in arbitrary units (**P = 0.0085, n = 5 mice per group, Student's t‐test). Scale bar, 10 μm.
• C
  Immunofluorescence images against CB₁ in Purkinje cells of the cerebellum of age‐matched control and NPC‐affected children. Graph shows mean ± SEM intensity associated with CB₁ in the Purkinje cells expressed as percentage of the control values (16 and 57 cells in control and NPC, respectively). Scale bar, 10 μm.
• D
  Graph shows mean ± SEM SM levels, expressed as percentage of vehicle values, in cultured fibroblasts from a NPC patient treated with vehicle or with PF at 50 or 100 μM (n = 2 different cultures)
• E
  Graph shows mean ± SEM cholesterol levels, expressed as percentage of vehicle values, in cultured fibroblasts from a NPC patient treated with vehicle or with PF at 50 or 100 μM (n = 2 different cultures)
• F
  Immunofluorescence images of CB₁ and Lamp1 in cultured fibroblasts from control subject and NPC patient treated with vehicle or with PF at 100 μM. Graph shows mean ± SEM Mander's coefficient for the co‐localization of CB₁ with Lamp1 (n = 2 different cultures). Scale bar, 5 μm.
• G
  Graphs show mean ± SEM SM and cholesterol levels expressed as percentage of the values obtained in the vehicle‐treated mice in the cerebellum of Npc1^nmf164 after 48 h of a single dose of 5 mg/kg PF (n = 3 mice)
• H
  Immunofluorescence images against the microglia marker F4/80 in the cerebellum of WT and Npc1^nmf164 after 48 h of a single dose of 5 mg/kg PF. Graphs show mean ± SEM number (upper) and area (lower) of F4/80-positive cells (upper graph: ****P < 0.0001; lower graph: ***P _{WT Veh vs NPC Veh} = 0.0008; *P _{WT Veh vs NPC PF} = 0.0286; ****P _{WT PF vs NPC Veh} < 0.0001; **P _{WT PF vs NPC PF} = 0.0022, n = 3 mice per group, > 20 cells analyzed per mouse, one‐way ANOVA, Bonferroni post hoc). Scale bar, 10 μm.

Source data are available online for this figure.