Fig 1: High TBX1 expression is related to poor prognosis in PCa. (A) Left, TBX1 is expressed at high levels in PCa tissues, and TBX1-positive cells exhibit nuclear and cytoplasm staining. Right, TBX1 is absent in non-neoplastic tissues. (B) Kaplan–Meier curves demonstrating the survival distribution stratified by TBX1 expression level. Left, biochemical recurrence-free survival determined from 280 PCa samples in the study. Right, disease/progression-free survival determined from 499 PCa samples in the TCGA database. (C) Analysis of microarray TBX1 expression data from the Oncomine database. Analysis of the log2 median intensity values of TBX1 was conducted using six different datasets. TBX1 is commonly expressed at high levels in PCa tissues. *p < 0.05; **p < 0.01; ***p < 0.001. Prostate cancer, PCa; Non-neoplastic tissue adjacent to the tumor, NTAT; Biochemical recurrence (BCR).
Fig 2: Analysis of significantly enriched GO annotations and KEGG pathways of differentially expressed genes correlated with TBX1 from PCa patients in the TCGA database using GSEA. The blue column represents the normalized enrichment scores. FDR < 0.05. Gene Ontology, GO; Gene Set Enrichment Analysis, GSEA.
Fig 3: TGR5 activation induces scWAT beiging in mice fed a high-fat diet. a Body weight curves of TGR5 wild-type (Tgr5+/+) fed a high-fat (HF) diet for 20 weeks in the presence or absence of the selective TGR5 agonist INT-777. n = 10 per group. b mRNA levels of beige remodelling markers Pgc1a, Ucp1, Tbx1, Prdm16, Cidea, Cd137, Pparg2 and Cebpb in the scWAT of mice described in a. c Representative (n = 10 per group) western blot of PGC-1α, the mitochondrial marker VDAC1, and beiging markers TBX1 and UCP1 from the scWAT of mice described in a. GAPDH was used as loading control. d Quantification of mitochondrial (16S) vs. nuclear (HK2) DNA ratio from the scWAT of mice described in a. e Representative haematoxylin and eosin staining from the scWAT of mice described in a. f Adipocyte area quantification from images shown in e. g Representative (n = 5 per group) UCP1 immunostaining of scWAT sections from mice described in a. h Quantification of UCP1 immunostaining intensity depicted in g. Scale bars = 50 μm. Results represent mean ± SEM. *P ≤ 0.05 and **P ≤ 0.01 vs. Tgr5+/+ HF group by two-way ANOVA followed by Bonferroni post hoc test (a) or Student’s t test (b, d, h). Uncropped western blots are provided in Supplementary Fig. 12A and B
Fig 4: H3K4me1 enhancement partially counteracts the effect of TBX1 silencing. (A) Left, Western blot results showing that TCP treatment partially rescues the decrease in H3K4me1 due to TBX1 silencing. Right, quantitative results of the left panel. (B) CHIP–qPCR analysis results showing that TCP treatment partially rescues the observed decrease in level of H3K4me1 binding with rRNA genes due to TBX1 silencing. (C) CHIP–qPCR analysis results showing that TCP treatment partially rescues the decrease in level of UBF binding with rRNA genes due to TBX1 silencing. (D) qPCR analysis results showing that TCP treatment partially rescues the decrease in 45S rRNA due to TBX1 silencing. (E) TCP treatment partially rescues the decrease in colony formation due to TBX1 silencing. Upper panel, representative images of colony formation. Lower panel, quantitative analysis of colon numbers. The data represent the mean ± SD of three independent experiments. *p < 0.05; **p < 0.01, shTBX1 vs. shTBX1+TCP/shCtrl. Tranylcypromine, TCP; upstream binding factor, UBF.
Fig 5: TGR5 activation promotes beige adipocyte differentiation in vitro. a Heat map showing the expression of Tgr5 and adipocyte and beiging markers from human bone marrow-derived mesenchymal stem cells induced for adipogenic differentiation (GEO Accession number GSE80614)32. Colour key represents row z-score. b mRNA levels of beige remodelling markers Pgc1a, Ucp1, Tbx1, Prdm16, Cidea, Cd137, Pparg2 and Cebpb assessed in the human pre-adipocyte cell line, Simpson Golabi Behmel Syndrome (SGBS). SGBS cells were differentiated in presence or absence of the TGR5 agonist INT-777. n = 6. c mRNA levels of genes described in b in adipocytes differentiated from the stromal vascular fraction (SVF) of TGR5 wild-type (Tgr5+/+) and germline TGR5 knock-out (Tgr5−/−) mice. SVF cells were differentiated for 7 days in presence or absence of the TGR5 agonist INT-777. n = 6. d Representative (n = 6 per group) western blot of PGC-1α, mitochondrial markers (VDAC1 and TOMM40) and beiging markers TBX1 and UCP1 from the cells described in c. PARP1 was used as loading control. e Quantitative densitometry of the western blots showed in d. f Quantification of mitochondrial (16S) vs. nuclear (HK2) DNA ratio from the cells described in c. g Spare respiratory capacity of the cells described in c, calculated as the difference between maximal (FCCP) and basal oxygen consumption rate (OCR). h, i Glycerol (h) and fatty acid (i) release from the cells described in c after 1 h stimulation with the TGR5 agonist INT-777 or vehicle (DMSO). j Basal oxygen consumption rate (OCR) of cells described in c after 3 h pre-incubation with etomoxir and stimulation with the TGR5 agonist INT-777 or vehicle (DMSO). Results represent mean ± SEM. *P ≤ 0.05, **P ≤ 0.01 and ***P ≤ 0.001 vs. Tgr5+/+ cells by one-way ANOVA followed by Bonferroni post hoc test (c, e–j) or Student’s t test (b). Uncropped western blots are provided in Supplementary Fig. 13A–C and Supplementary Fig. 14A
Supplier Page from Abcam for Anti-TBX1 antibody [EPR3289(2)]