Figure 7.

c-Abl Deficiency Reduces Cholesterol Accumulation in NPC In Vivo Models

(A) Representative immunofluorescence showing cholesterol by GST-PFO staining (red) in cerebellum sections from 8-month-old wild-type (WT) and NPC mice treated with vehicle or imatinib 12.5 mg/Kg, every day for 2 weeks by intraperitoneal injection. Arrows show purkinje neurons. Scale bars, 50 μM. (B) Quantification of cholesterol accumulation per Purkinje cell area in (A) (n = 2) (5 images per animal).

(C–E) WT and NPC mice were treated for 4 weeks with vehicle and GNF-2 (5 mg/kg in 40% water, 30% polyethylene glycol 300 and 30% propylene glycol) starting at p28. Motor coordination was assessed weekly by beam test (C), weight was registered during the treatment (D) and (E) Cerebella from vehicle and GNF-2 treated female NPC mice were analyzed at 8 weeks of age for calbindin by immunohistochemistry. Quantification of Purkinje cell area/Purkinje cell layer (PCL) area is shown (n = 9). Scale bars, 100 μM.

(F) WT and NPC mice were treated with GNF-2 (5 mg/Kg) for three weeks starting at p28. Upper panels show filipin staining of cerebellum sections and lower panels show filipin staining of liver sections. n = 3 independent experiments. Scale bars, 50 μM.

(G) Representative immunofluorescence showing cholesterol by GST-PFO (red) and endogenous TFEB (green) in cerebellum sections from 8-month-old wild-type (WT) and c-Abl KO mice treated with vehicle or U18666A 10 mg/Kg, for 2 days by intraperitoneal injections. Arrows show purkinje neurons. Scale bars, 50 μM. Statistical analysis with one-way ANOVA followed by Tukey's post-test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Data represent mean ± SEM.