Supplemental Materials

Metabolomics analysis identified lipid and bile acid markers linked to anti-diabetic and

anti-obesity function of bile acids in mice

Yunpeng Qi, Changtao Jiang, Jie Cheng, Kristopher W. Krausz, Tiangang Li, J. M. Ferrel, Frank

J. Gonzalez and John Y.L. Chiang

Fig. S1. A typical chromatogram generated using the lipidomics method presented in this study.

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1.4813.03

0.78

0.64

0 18.0016.0014.0012.0010.008.006.004.002.00 Time

%

100

Fig. S2. PCA analysis of CYP7A1-tg and WT mice challenged with HFD shows distinct lipidomics profiles of these two groups. Cumulative R2X 0.677 for serum (A) and 0.593 for

liver (B). Abbreviations: tg, transgenic mice overexpressing CYP7A1 in the liver; wt, wild-type

mice.

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Fig. S3. Identification of a significant marker ([M+H]+ m/z 782.5695). (A) Extracted ion

chromatogram (EIC) of m/z 782.5695 from standard substance (Rt=9.56min); (B) EIC of m/z

782.5695 from the sample (Rt=9.51 min); (C) MS/MS spectrum of the commercial standard of

PC 16:0-20:4; (D) MS/MS spectrum of the ion in the sample.

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Fig. S4. Lipid marker quantitation results in ileum and liver by multiple reaction-monitoring mass spectrometry based on standard curves using authentic standards. CYP7A1-tg mice had declined levels for the detected lipid markers in ileum (A) but not in liver

(B). Data were expressed as mean ± SD. Significant comparison was based on two tailed

Student’s t-test or Mann-Whitney test. An *indicates p < 0.05 and an **indicates p<0.01 (with

respect to the WT group). Abbreviations: WT-CTR, control diet-treated wild-type mice; Tg-CTR,

control diet-treated CYP7A1 transgenic mice.

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Fig. S5. PCA analysis of the data from mouse gallbladder, liver, ileum and colon samples reveal distinct metabolic profiles of the CYP7A1-tg and WT mice. Cumulative

R2X 0.923 for gallbladder (A), 0.659 for liver (B), 0.939 for ileum (C) and 0.708 for colon (D).

Abbreviations: Tg, transgenic mice overexpressing CYP7A1 in the liver; WT, wild-type mice.

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Fig. S6. Heat map of bile acid marker levels in the ileum of WT and CYP7A1-tg groups. Rows: sample; columns: bile acids. Color key indicates metabolite expression value, green:

lowest, red: highest. Abbreviations: Tg, transgenic mice overexpressing CYP7A1 in the liver;

WT, wild-type mice.

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