Fig 1: Dysregulated BA profiles in the HF-OP group and relative expansion of non-12-OH BAs composition in the HF-OR group. (a) Orthogonal partial least squared-discriminant analysis (OPLS-DA) scores plot of serum BA profiles showing the groupings of N (blue) group, HF-OP (red) and HF-OR (green). (b) VIP scores of OPLS-DA based on the serum BA profiles between the HF-OP and HF-OR group. A BA with VIP more than 1 was considered important in the discrimination between the groups. (c) Dysregulated BAs in the HF-OP group. The data are presented as the mean ± SD. *p<0•05 and **p<0•01 (Mann–Whitney U test). (d) Heatmap of spearman correlation coefficients between serum BAs and blood biochemical parameters from all samples in the three groups (n = 24, 8 samples per group). The gradient colours represent the correlation coefficients, with red color being more positive and blue color indicating more negative. *p<0•05 (Spearman's correlation with the post hoc correction using the Holm method). (e) Differential BAs in liver tissue with a relative expansion of non-12-OH BA composition in HF-OR group. The data are presented as the mean ± SD. *p<0•05 and **p<0•01 (Mann–Whitney U test), ns showed no significance. (f) Pie graphs show the mean percentage of non-12-OH BAs in the liver among the three groups. (g) The relative mRNA levels of Fxr, Shp and critical enzymes responsible for BA synthesis in liver among the three groups. The data are presented as the mean ± SD. **p<0•01 (unpaired Student's t-test). n = 5 per group. (h) The expression of CYP8B1 (n = 4 per group) and CYP7B1 (n = 4 per group) in liver were detected by western blot. Data are presented as the mean ± SD. *p<0•05 and **p<0•01 (unpaired Student's t-test). (i) The expression of FXR and FGF15 in ileum were detected by western blot. Data are presented as the mean ± SD. *p<0•05 (unpaired Student's t-test). (j) Serum FGF15 levels were detected by ELISA. Data are presented as the mean ± SD. **p<0•01 (unpaired Student's t-test).
Fig 2: Expression of endocrine FGFs and FLRT2 in plasma. a Comparison of FGF15, FGF21, FGF23 and FLRT2 expression levels in response to fasting in male mice plasma. b Comparison of FGF15, FGF21, FGF23 and FLRT2 expression levels in response to fasting in female mice plasma. c Comparison of FGF15, FGF21, FGF23 and FLRT2 expression levels in female and male mice plasma. Data represent means ± S.E.M, n = 6; biological replicates. *P < 0.05, **P < 0.01 and ***P < 0.001 by one-way ANOVA with unpaired two-tailed Student’s t test
Fig 3: Effects of whey protein isolate (5 g/100 ml water) or ß-conglycinin (5 g/100 ml water) intake on expression of FXR (A, D), FGF15 (B, E), ASBT (G, I), Osta, and Ostß (H, J) in the ileum, and plasma FGF15 levels (C, F) in mice. The relative amount of mRNA is shown as fold-change of the mean value of the control group in C57BL6J mice fed a chow. Data are presented as the mean ± SEM (n = 6/group). *P < 0.05.
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