Statistical tests were performed using Prism version 7.0b for Mac OS X (GraphPad Software, La Jolla California USA) or using R packages ( To assess weight loss a two-way ANOVA with Dunnett’s multiple comparisons post hoc test comparing ursodiol treatment groups and untreated mice was performed. For microbiome analysis, analysis of molecular variance (AMOVA) was used to detect significant microbial community clustering of treatment groups in NMDS plots and principle coordinate analysis (PCoA) biplots using Spearman correlation were used to examine difference in microbial community structures between ursodiol treatments and compared to pretreatment [41]. For bile acid metabolome, a NMDS illustrates dissimilarity indices via Horn distances between bile acid profiles. To assess the comprehensive bile acid profiles, a two-way ANOVA followed by Dunnett’s multiple comparisons post hoc test was used to compare ursodiol treatment groups to pretreatment bile acid profiles. A Kruskal-Wallis one-way ANOVA test followed by Dunn’s multiple comparisons test was used to calculate the significant of individual bile acid within each ursodiol treatment group compared to pretreatment. Statistical significance was set at a p value of < 0.05 for all analyses (*, p <0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001).

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