Fig. 4. Abnormalities in adult zebrafish visceral organs. A: Lipid levels were determined in pmol/mg liver tissue. GlcSph, GalCer, and GlcChol were separated from their respective galactosylated lipid by HILIC chromatography. Data is depicted as mean ± SEM; WT (n = 10), gba1-/- (n = 9), asah1b-/- (n = 8), and gba1-/-:asah1b-/- (n = 11). Data is analyzed by One-Way Anova with Tukey’s multiple comparison test. B: Expression of gpnmb, chia.6, il-1β, tnfβ, apoEb, or catD mRNA levels were determined using RT-qPCR analysis; WT (n = 6–9), gba1-/- (n = 6–8), asah1b-/- (n = 5), and gba1-/-:asah1b-/- (n = 7). Data is normalized using two housekeeping genes (ef1α and rpl13) and analyzed by One-Way Anova with Tukey’s multiple comparison test. Statistical analysis is depicted of WT versus gba1-/-, asah1b-/-, or asah1b-/-:gba1-/-, only when a significant difference is apparent or of gba1-/- versus asah1b:gba1-/- when significant difference is of interest. Ns = not significant, ∗p < 0.05,∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001. C–E, Zebrafish were serially sectioned along the sagittal plane and stained using H&E. Images of liver, spleen, and pancreatic tissue of WT (C), gba1-/- (D), and gba1-/-:asah1b-/- (E) zebrafish. Areas of interest are marked by dotted boxes, and higher magnifications of these areas are shown below the respective overview images. Scale bars: 50 μm and 20 μm for low and higher magnifications, respectively. GlcSph, glucosylsphingosine; GalCer, galactosylceramide; GlcChol, glucosylated cholesterol.
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