Fig. 5

5A-SM mobilizes cholesterol in vivo and ameliorates disease phenotypes. a Serum cholesterol from 7-week-old Npc1 I1061T homozygous mice pre- and 2 h post-treatment with 100 mg/kg 5A-SM, i.p. b Pre- (dashed line) and 2 h post-treatment (solid line) serum was fractionated by HPLC, and cholesterol was quantified by cholesterol oxidase assay. VLDL, LDL, and HDL fractions are indicated by arrows. c, d Seven-week-old wild-type (WT) and Npc1 I1061T homozygous (NPC) mice were injected i.p. with vehicle (Veh) or 100 mg/kg 5A-SM. 48 h later, c liver HMGCS transcript levels and d total serum bilirubin were analyzed. e WT and Npc1 I1061T mice were injected i.p. with vehicle (Veh) or 100 mg/kg 5A-SM three times per week from 7 to 11 weeks of age. The change in weight of each mouse from week 7 (t = 0) to week 11 (t = 4) was quantified. f Seven-week-old WT and NPC mice were injected with Veh or 100 mg/kg 5A-SM three times per week for 2 weeks. At 9 weeks of age livers were stained for macrophages using F4/80 (green) and DNA Hoechst (blue). Macrophage area is quantified at right. Scale bar = 50 μm. Violin plot shows median (dashed line), 25% and 75% (dotted lines), and probability density (thickness). g Brain slices from 8-week-old Npc1 I1061T mice were incubated with vehicle (Veh) or 5 mg/ml 5A-SM for 4 days, and filipin levels in Purkinje neuron soma were quantified (see also Additional file 1: Figure S4b). h Six- to 7-week-old WT and Npc1 I1061T mice received intraventricular injections with vehicle (Veh) or 5A-SM-DiD. N: WT = 4, NPC Veh = 5, NPC 5A-SM = 4 mice. One-week later, cholesterol levels in Purkinje neuron soma (green) were analyzed by filipin (blue) staining. Dashed lines indicate Purkinje neuron soma (also see Additional file 1: Figure S6a). Scale bar = 50 μm. Data quantified at right. Data are mean ± s.e.m. from a, b, c three; d genotype and treatment: number of mice, WT + Veh = 5, WT + 5A-SM = 3, NPC + Veh = 4, NPC + 5A-SM = 7; e genotype and treatment: number of mice at 9 weeks and 11 weeks, WT + Veh: 13 and 8, WT + 5A-SM: 9 and 8, NPC + Veh: 6, NPC + 5A-SM: 12 and 10 mice; f genotype and treatment: number of mice, cells, WT + Veh: 4, 301, NPC + Veh: 4, 514, NPC + 5A-SM: 3, 373 (g) WT = 93, NPC Veh = 143, NPC + 5A-SM = 116 cells. *p ≤ .05, **p ≤ .01, ***p ≤ .001, ****p ≤ .0001. a, c Student’s t test (t = (a) 6.375, (c) 5.23); d, f, g, h one-way ANOVA with Tukey post hoc test (F, df = (d) 13.28, 3; (f) 368.1, 2 (g) 38.89, 2; (h) 108.3, 2); e two-way ANOVA with Bonferroni post hoc test (F, df = 7.12, 2)