FIG 3.

Hepatic ganglioside abundance in miglustat-treated mice. (A) Ganglioside synthetic pathways. Each step in the pathway is catalyzed by a specific enzyme (39). GCS, glucosylceramide synthase, encoded by the gene UGCG. (B) Class composition of gangliosides in untreated Ifnar1^−/− mouse livers (n = 8) (left) versus Huh-7.5 cells (n = 3 technical replicates) (right). Huh-7.5 cell data are from a study by Das et al. (13). (C) Ion intensities of different ganglioside classes in liver tissues from miglustat-treated and untreated naive mice (n = 1 each) (left) and HAV-infected mice (n = 2 and 3, respectively) (right) in the experiment shown in Fig. 2. Based on summed intensities, miglustat reduced the overall ganglioside abundance in HAV-infected livers by 74.9%. GM3 ganglioside was reduced by 73.7%, GM2 was reduced by 82.7%, GM1 was reduced by 78.8%, and GD1a was reduced by 78.3% compared to untreated mice. GD3 comprised less than 0.01% of gangliosides in both treated and untreated animals. (D) Class composition of gangliosides in livers from untreated and miglustat-treated mice. Data shown are the mean percentages that each ganglioside class represented of all gangliosides, ± SD, based on summed intensities. Each symbol represents the liver from an individual animal. (E) Volcano plot showing differences (fold changes) in the abundances of 56 individual ganglioside species in a repeat analysis of the liver tissues from treated versus untreated HAV-infected mice (see Table S1 in the supplemental material). Probability was determined by an unpaired t test with Šidák-Holm correction for multiple comparisons. Solid symbols indicate a false discovery rate (FDR) of <0.01%; the horizontal line indicates a P value of 0.05.