Fig 1: Liver and brain damage in AOM-induced acute liver failure model mice. (A) H&E staining of normal mouse liver, liver section from a preterminal stage IV HE mouse 35 h after exposure to AOM, liver section from a mouse 35 h after exposure to AOM and previous administration of budesonide or SC-435 for 5 days (Magnif. 200×)). (B) ALT levels in C57BL/6 J mice with HE due to AOM-induced liver failure. (C) H&E staining of a brain section of a healthy control mouse, brain section of a mouse exposed to AOM for 35 h (stage IV HE), AOM + SC-435 mice and AOM-budesonide mice showing astrocyte ballooning and twinning (arrow). Bar scale: 73 µm. (D) Immunohistochemistry for GFAP on a brain section from a normal mouse, in stage IV HE AOM mice, AOM + SC-435 mice and AOM-budesonide mice. (E) GFAP positive cell percentage in three groups. (F) Immunohistochemistry for NeuN on a brain section from a normal mouse, in stage IV HE AOM mice, AOM + SC-435 mice and AOM-budesonide mice. (G) NeuN positive cell percentage in three groups. (H) Time (s) spent in the open arms of the maze at different groups. (I) Survival rates of mice treated with vehicle, AOM, AOM + SC-435, and AOM + Budesonide. (J, K, L, M) Total BAs in serum, liver and brain were significantly increased while total BAs in ileum content were significantly decreased in AOM-induced model mice and were further increased by budesonide treatment but were normalized after the SC-435 intervention. (N) Ileum content pH of the 4 study groups. (O) Serum ammonia levels of the 4 study groups. (P) Liver FXR levels. (Q, R, S, T) Ileum and brain BA transporters were significantly increased in AOM-induced model mice and were further increased by budesonide treatment but were normalized after the SC-435 intervention. a, p < .05, compared to normal group; b, p < .05, compared to AOM group; c, p < .05, compared to AOM/Budesonide group.
Fig 2: FXR and PD-L1 expression are associated with tumor response to anti-PD-1-based chemo-immunotherapy in patients with NSCLC. (A) Representative IHC images of FXR (upper panel) and PD-L1 (lower panel) expression in NSCLC specimens (Scale bar, 50 µm). Isotype control: the primary antibody was replaced by nonspecific mouse or rabbit IgG. (B and C) Representative IHC images (upper graphs) and IHC scores (lower graphs) of FXR (B) and PD-L1 (C) in responders vs. non-responders (P=0.01 and P=0.003, respectively; Mann-Whitney U test). Scale bars indicate 50 µm. Error bars indicate the median and interquartile range. (D) Objective response in patients with low FXR vs. high FXR (ORR 22.5 vs. 38.5%, P=0.036; chi-square test). (E) Objective response in patients with low PD-L1 vs. high PD-L1 (ORR 22.1 vs. 46.3%, P=0.002; chi-square test). The objective response rate (n/N), is shown above each bar. FXR, farnesoid X receptor; PD-L1, programmed death-ligand 1; PD-1, programmed death-1; NSCLC, non-small cell lung cancer; IHC, immunohistochemistry.
Fig 3: Regulatory effects of unconjugated and conjugated BAs on ASBT and FXR protein expression. (A) Proportion of different BAs in intestinal content in AOM mice and normal controls. FC was calculated as the ratio AOM/normal of the average proportion of BAs. (B) Western-blot analysis of the expression of ASBT in human cecum epithelial cell line CLL-251 treated with 5 groups of BAs (5 unconjugated BAs and their taurine and glycine conjugates, 50 µM, 48 h). Beta-actin was used as loading control. (C, D, E) The expression of ASBT and FXR in the CLL-251 3D micro-tissues with different indicated treatments detected by immunofluorescent staining. (F) Immunofluorescent staining of mouse intestine tissues in normal and AOM mice for the expression of ASBT and FXR. The mean of florescence Intensity (MFI) of ASBT and FXR nuclear positive cells percentage were calculated by Image J.
Fig 4: FXR knockdown activates the Wnt signaling pathway. (A) FXR and ß-catenin protein expression levels detected by western blotting in the HT-29, Caco-2 and HCT-116 cell lines. ß-actin was used as a loading control. (B) FXR silencing was confirmed in all cell lines. (C) Silencing FXR significantly increased ß-catenin-mediated luciferase activity. The error bars represent the SEM of three independent samples. *P<0.05 vs. siNC. FXR, farnesoid X receptor; si, small interfering; NC, negative control.
Fig 5: Subgroup analysis of tumor responses and prognosis based on the IHC levels of FXR and PD-L1 in patients with NSCLC receiving anti-PD-1-based chemo-immunotherapy. (A) Objective response in patients with FXRhighPD-L1high, FXRlowPD-L1high, FXRhighPD-L1low, or FXRlowPD-L1low (ORR in FXRhighPD-L1low vs. FXRlowPD-L1low patients, 31 vs. 8.1%, P=0.009; chi-square test). The objective response rate (n/N), is shown above each bar. (B) Representative IHC images (upper panels) and IHC score (lower panel) of FXR in PD-L1low responders vs. PD-L1low non-responders (P<0.001; Mann-Whitney U test). Scale bars indicate 50 µm. Error bars indicate the median and interquartile range. (C and D) Kaplan-Meier survival curves for PFS and OS of four subgroups (PFS and OS in FXRhighPD-L1low vs. FXRlowPD-L1low patients, P=0.013 and P=0.03, respectively; log-rank test). IHC, immunohistochemistry; FXR, farnesoid X receptor; PD-L1, programmed death-ligand 1; NSCLC, non-small cell lung cancer; PD-1, programmed death-1; PFS, progression-free survival; OS, overall survival.
Supplier Page from Abcam for Anti-Bile Acid Receptor NR1H4 antibody [322.1.2.2] - N-terminal