Article Figures & Data
Figures
- Fig. 1
Relative hepatic and ileal mRNA values of Fgf15-related genes and RNA-seq analysis. (A) Hepatic and ileal gene expression was normalized to β-actin mRNA expression and graphs depict relative mRNA ± 1 S.D. (n: WT = 9, Fgf15int−/− = 6, Fgf15 Tg = 5; one-way ANOVA, Tukey post hoc). An asterisk denotes a significant difference from WT (P < 0.05). (B) Heat map illustrates logtwofold-change in RNAseq analysis as compared with WT (n = 3/group).
- Fig. 2
Relative hepatic mRNA, protein, and function of cytochrome P450s. (A) Relative hepatic mRNA expression normalized to β-actin mRNA expression (n: WT = 5, Fgf15int−/− = 6, Fgf15 Tg = 5; one-way ANOVA, Tukey post hoc). Graphs depict relative mRNA ± 1 S.D. An asterisk denotes a significant difference from WT (P < 0.05). Cyp2b9 and Cyp2b10 failed Levene’s test, therefore Kruskal-Wallis was used for analysis. (B) Western blot of CYP3A and CYP2B using 20μg of liver homogenate (n: WT = 4, Fgf15int−/− = 5, Fgf15 Tg = 5; one-way ANOVA, Tukey post hoc). (C) Liquid chromatograhy-mass spectrometry analysis of relative protein expression of CYP2B and CYP3A isoforms (n: WT = 5, Fgf15int−/− = 5, Fgf15 Tg = 5, WT-PB = 3; one-way ANOVA, Tukey post hoc). (D) Measurements of pentoxyresorufin and midazolam metabolite formation to assess CYP2B and CYP3A activity in liver microsomes of WT, Fgf15int−/−, Fgf15 Tg, and phenobarbital-treated WT mice (n: WT = 5, Fgf15int−/− = 5, Fgf15 Tg = 5, WT-PB = 3; one-way ANOVA, Tukey post hoc).
- Fig. 3
Role of CAR in alterations brought about by the overexpression of Fgf15/19. (A) Fluorescent imaging of Ad/EYFP-hCAR PHHs treated with PB, CITCO, or FGF19 recombinant protein. (B-E) Hepatic gene expression was normalized to β-actin mRNA expression, and graphs depict relative mRNA ± 1 S.D. An asterisk denotes a significant difference from WT (P < 0.05). Cyp2b10, Cyp2b9, Cyp2b13, and Cyp17a1 failed Levene’s test, therefore Kruskal-Wallis was used for analysis. (B) Relative hepatic mRNA expression of genes regulated by CAR. (C) Relative hepatic mRNA expression of genes involved in drug metabolism. (D) Relative hepatic mRNA expression of genes predominantly expressed in male mice. (E) Relative hepatic mRNA expression of genes predominantly expressed in female mice (n: WT = 6, Fgf15 Tg = 7, CAR−/− = 6, Fgf15 Tg/CAR−/− = 7; one-way ANOVA, Tukey post hoc).
- Fig. 4
Impact of Fgf15 overexpression on the pharmacokinetics of bupropion in mouse plasma. (A) Average log concentrations of bupropion and hydroxybupropion (+S.D.) in WT and Fgf15 Tg mouse plasma. (B) Table summarizing pharmacokinetic parameters (±S.D.) of bupropion and hydroxybupropion in wild type and Fgf15 Tg mouse plasma (n: WT = 10, Fgf15 Tg = 8; Student’s t test).
- Fig. 5
Body size and composition of WT and Fgf15 Tg mice. (A) Representative dorsal view of WT (top) and Fgf15 Tg (bottom) mice. (B) Quantification of nose to anus length (cm) and body weight (g) of WT and Fgf15 Tg mice (n: WT = 5, Fgf15 Tg = 6; Student’s t test). (C) Percent of total body weight distributed as fat, muscle, and bone (n: WT = 5, Fgf15 Tg = 5; Student’s t test). (D) Western blots and semiquantifications of cell signaling proteins in liver homogenates from WT and Fgf15 Tg mice (n: WT = 4, Fgf15 Tg = 4). (E) Relative hepatic gene expression of GH response genes Igf-1 and Igfals (n: WT = 6, Fgf15 Tg = 7; Student’s t test). (F) Serum GH (left) and IGF-1 (right) levels in WT and Fgf15 Tg mice measured by ELISA every 30 minutes from 9:30 AM to 2:30 PM.
Additional Files
Data Supplement
- Supplemental Data -
Supplemental Fig. 1. Primer, antibody, and animal information.
Supplemental Fig. 2. In vitro assays showing mRNA expression of selected genes following treatment with recombinant FGF19 protein.
Supplemental Fig. 3. Liver histology and function assessment.
Supplemental Fig. 4. Generation of Fgf15 Conditional Knockout Mice.
Supplemental Fig. 5. Serum (A) GH and (B) IGF-1 levels for female WT and Fgf15 Tg
Supplemental Table 1. Surrogate peptides used for LC-MS/MS-based protein quantification.
Supplemental Methods.
Supplemental Methods Table 1. Acquisition parameters use in the LC-MS/MS analysis.
- Supplemental Data -
In this issue
Effects of Overexpression of FGF15/19 on Hepatic DMEs
