Fig 1: Effect of TPE-CA on FXR-targeted protein. (A) The level of FXR in liver for four groups. (B) The level of BSEP for four groups. (C) The level of NTCP for four groups. (D) The level of OATPs for four groups. Expressions are normalized to GAPDH. **p < 0.01, ***p < 0.001. Values are represented as means ± S.D for 10 animals per group.
Fig 2: Characterization of HepG2/C3A spheroids after 21 days of dynamic culture in a biochip containing the hydroscaffold. (a) F-actin and MRP2 staining showing the formation of bile canalicular-like structures: DAPI (nuclei, blue), phalloidin (F-actin, green); MRP2 (red) and biliary-like network (co-localization MRP2 and F-actin signals, yellow overlay signal, the two pictures in the bottom correspond to an enlargement from the merge picture); (b) BSEP staining: DAPI (nuclei, blue) and BSEP (yellow); (c,d) albumin and urea production in the dynamic biochip and static Petri conditions throughout 21 days of culture (both biochip and Petri contained the hydroscaffold). The immunostaining images (a,b) correspond to z-stack projections. * p < 0.05.
Fig 3: SGPA activating osteoblast FXR-RUNX2 signalling in vivo. (A) The mRNA expression of Bsep and Runx2 in bone tissues of OVX-induced osteoporosis mice were analysed by qRT-PCR (n = 6). (B–C) The representative western blotting analysis and relative quantification of BSEP and RUNX2 protein levels in bone tissues of OVX-induced osteoporosis mice (n = 3). (D) Immunostaining of FXR (green) and Runx2 (red) on femur tissue of OVX-induced osteoporosis (magnification, 630 × ). (E) The relative quantification of FXR and Runx2 levels in femur (n = 3). Data are presented as the means ± S.D. n.s not significant, ∗P < 0.05, ∗∗P < 0.01.
Fig 4: (A) The relative luciferase activities of six candidate compounds were evaluated at a single concentration of 20 µM (B) The relative luciferase activities of compound 2 in a concentration-dependent manner (*p < 0.05, **p < 0.01, ***p < 0.001 vs control group; data is expressed as mean ± SD.). (C) Western blot analysis for FXR, SHP and BSEP expressions after treatment 24 h in HepG2 cells (n = 3, data is shown as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 vs. control group.).
Fig 5: CDCA content and inflammasome expression were both suppressed in MRP2-/- mice. (A) High and low concentrations of BAs in liver tissues of MRP2−/− mice (n = 5) and WT mice (n = 4) were detected by LC-MS (**p < 0.01 and ***p < 0.001). (B) The relative ratio of GCDCA and TCDCA to CDCA concentrations in MRP2−/− mice (n = 5) and WT mice (n = 4) determined by LC-MS. (p < 0.05 compared with WT). (C) Western blot analysis of inflammasome Pro-caspase-1 cleavage in liver tissues of MRP2−/− mice (n = 5) and WT mice (n = 4); β-actin was used as a loading control, and the band intensities were quantified by Image J software and shown as a bar graph. Quantification of the image was shown as means ± SD, and **p < 0.01 compared with WT. (D) Heat map analysis of the mRNA of 20 genes that control BA homeostasis in MRP2−/− mice (n = 3) and WT mice (n = 3) detected by qPCR. (E) The protein expression of FXRα, Cyp8b1, Cyp27a1, Sult2a1, Bsep, Mrp3, Ntcp, and BAAT in liver tissues of WT (n = 4) versus MRP2−/− mice (n = 5). GAPDH was used as a loading control, and the band intensities were quantified by ImageJ software and shown as a bar graph. Quantification of the image was shown as means ± SD, *p < 0.05, and ***p < 0.001 compared with WT. Abbreviations: BAs, bile acids; CDCA, chenodeoxycholic acid; FXR, farnesoid X receptor; TCDCA, taurochenodeoxycholic acid; WT, wild type.
Supplier Page from Abcam for Anti-ABCB11/BSEP antibody