Article Figures & Data
Figures
- Fig. 1.
Body weight and liver function. For each experimental group, body weight (A), serum AST (B), and ALT (C) activity were assessed at the end of the experiment. One-way ANOVA, alpha = 0.001. n = 6 mice/group. ctrl, control.
- Fig. 2.
Differential mRNA expression pattern. Volcano plot (A) and pathway enrichment analysis (B) comparing liver transcriptomes of VPA and VPA + OCA groups. n = 6 mice/group. Real-time RT-PCR of the genuine FXR target genes Shp (C) and for the organic solute transporter β (Ostbeta) (D). One-way ANOVA, alpha = 0.001, followed by Tukey’s post hoc analysis. n = 6 mice/group. ctrl, control.
- Fig. 3.
Hepatic oxidative stress in vivo. NGS-derived heat-map, n = 6 mice/group (A). Real-time RT-PCR of Gpx1, Gpx2, and Gpx6 and the glutathione transferase (Gst) a2. One-way ANOVA, alpha = 0.001, followed by Tukey’s post hoc analysis. n = 6 mice/group. (B–E) Representative staining of 4-HNE, Grp78, and glutamine synthetase staining (F). Scale bar, 100 µm. Adh4, alcohol dehydrogenase 4; Akr1b7, aldo-keto reductase 1b7; Cont, control; Gclm, glutamylcysteine synthetase; GS, glutamine synthetase; Nqo1, NAD(P)H quinone dehydrogenase 1.
- Fig. 4.
Oxidative stress in vitro. Cells were incubated for 24 hours with OCA (2 µM) followed by 24 hours with VPA (2 mM). Real-time RT-PCR of the human oxidative stress-related genes GPX1, glutamate-cysteine ligase regulatory subunit, NAD(P)H quinone dehydrogenase 1 (NQO1), and superoxide dismutase 2 (SOD2). Data are expressed as fold change to the housekeeping gene hypoxanthine phosphoribosyltransferase. One-way ANOVA, alpha = 0.005, followed by Tukey’s post hoc analysis. n = 3 independent experiments. (A–D) Total intracellular ROS level. Data are expressed as percentage of the untreated control. One-way ANOVA, alpha = 0.005, followed by Tukey’s post hoc analysis. n = 3 independent experiments (E). Cytosolic (F) and mitochondrial (G) ROS levels. Cells were exposed to H2O2 as positive control. Data are expressed as ratio between the fluorescent signals at the indicated excitation wavelengths. One-way ANOVA, alpha = 0.005, followed by Tukey’s post hoc analysis. n = 3 independent experiments. Representative cytosolic ROS staining. Scale bar, 50 µm (H). Cont, control; GCLM, glutamylcysteine synthetase.
- Fig. 5.
Hepatic Cyp450 expression in vivo. NGS-derived heat-map, n = 6 mice/group (A). Real-time RT-PCR of Cyp2c39 and Cyp2c44. One-way ANOVA, alpha = 0.001, followed by Tukey’s post hoc analysis. n = 6 mice/group (B and C). Representative Western blotting of Cyp2cs (D), n = 5 mice/group from the same experiment. Representative staining of Cyp2cs. Scale bar, 100 µm (E).
- Fig. 6.
Lipid accumulation in vivo. NGS-derived heat-map of PPARγ-related genes, n = 6 mice/group (A). Real-time RT-PCR of the acyl-CoA thioesterase 1 (Acot1) and the PPARγ. One-way ANOVA, alpha = 0.001, followed by Tukey’s post hoc analysis. n = 6 mice/group (B and C). Representative hematoxylin and eosin (D), BODIPY 493/503 (E), and macrophage marker MAC387 (F) staining, n = 6 mice/group. ctrl, control. Dgat, Diglyceride acyltransferase; Fabp, fatty-acid-binding protein; Slc, solute carrier; Srebf, sterol regulatory element-binding protein gene.
Additional Files
Data Supplement
- Supplemental Data -
Supplementary Table 1
Supplementary R Script. EdgeR Bioconductor: Comparison of OCA + VPA over VPA
Supplementary Figure 1 - Lipid accumulation in vitro.
- Supplemental Data -
