Fig. 8: FXR activation inhibits the canonical YAP-TEAD transcription activator activity. | Nature Communications

Fig. 8: FXR activation inhibits the canonical YAP-TEAD transcription activator activity.

From: Disrupting bile acid metabolism by suppressing Fxr causes hepatocellular carcinoma induced by YAP activation

Fig. 8

a Luciferase activity assay in Huh-7 cells, tracking the canonical YAP-TEAD transcription activator activity. 5 µM GW4064 treatment was 24 h, ****P < 0.0001. b Immunoprecipitation of HA-FXR in the nuclear-enriched fraction of Huh-7 cells. c Immunoprecipitation of HA-FXR in the nuclear-enriched fraction of Huh-7 cells, 8xGTII and 3xIR1 plasmids were co-transfected with FXR and TEAD4 plasmids, as indicated. d Immunoprecipitation of FXR, YAP and pan-TEAD in the nuclear-enriched fraction of Huh-7 cells. 5 µM GW4064 treatment was 24 h. e ChIP-qPCR analysis of TEAD4 and HDAC1 binding at the 8xGTII TBE sites in the Huh-7 cells. 8xGTII plasmid was co-transfected with GFP or Flag-YAP5SA plasmids, as indicated. 5 µM GW4064 treatment was 24 h, ****P < 0.0001. f Immunoprecipitation of indicated proteins in the nuclear-enriched fraction of liver tissues. g Representative images of liver from the indicated mice. h Liver/body-weight ratios, ****P < 0.0001. i Serum BA concentrations, ****P < 0.0001. j Serum ALT levels, ****P < 0.0001. k–m qRT-PCR analysis of indicated genes in the liver tissue, ****P < 0.0001. **P = 0.0012, ***P = 0.0009 (Ctgf, k); **P = 0.006, ***P = 0.0005 (Cyr61, k); **P = 0.007 (Klf6, k); ***P = 0.0002 (Fos, k); *P = 0.042, ***P = 0.0002, ***P = 0.0005 in sequence (Cyp7a1, i); *P = 0.038, *P = 0.018, ***P = 0.0001, **P = 0.005 in sequence (Cyp8b1, i); **P = 0.004, ***P = 0.0005, **P = 0.0017 in sequence (Cyp7b1, i); **P = 0.0035, ***P = 0.0006 (Cyp27a1, i); *P = 0.016 (Bsep, m). n, o Luciferase activity assay in primary mouse hepatocytes with indicated genotype, tracking the canonical YAP-Tead transcription activity (n), and Fxr transcription activity (o), ****P < 0.0001, ***P = 0.0001 (n). 5 µM GW4064 and 3 µM TDI treatment was 24 h. Scale bars: 1 cm (g). n = 3 independent experiments (a, e, n, o), n = 5 biological replicates in each group (h–j), n = 3 biological replicates in each group (k–m). Data are presented as Mean ± SD, p-values were determined by 1-way ANOVA with Sidak’s test (a, e, h–o). Similar results were obtained from three independent experiments (b–d, f). Source data are provided as a Source Data file.

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