Journal: Scientific Reports

Article Title: Hepatokine Fetuin B expression is regulated by leptin-STAT3 signalling and associated with leptin in obesity

doi: 10.1038/s41598-022-17000-w

Figure Lengend Snippet: Correlations between serum Fetuin B and obesity indices and serum leptin in obese adults. The correlations between serum Fetuin B and the body mass index (BMI) ( a ), waist–hip ratio (WHR) ( b ), body fat mass (BFM) ( c ) and serum leptin ( d ) were assessed in 215 obese Chinese adults (n = 215). Pearson correlation coefficient was applied for correlation analysis. P values < 0.05 were considered statistically significant.

Article Snippet: The levels of human serum leptin (#ab179884), Fetuin B (#ab240684) and mouse serum leptin (#ab100718) were determined by ELISA (Abcam, Shanghai, China) according to the manufacturers’ instructions.

Techniques:

Journal: Scientific Reports

Article Title: Hepatokine Fetuin B expression is regulated by leptin-STAT3 signalling and associated with leptin in obesity

doi: 10.1038/s41598-022-17000-w

Figure Lengend Snippet: Expression of Fetuin B in HFD-induced obese mice. Body weight ( a , b ), body fat % ( c , d ), and serum leptin levels ( e , f ) were compared in mice fed chow with mice fed a HFD for 12 and 18 weeks (n = 5–12). The serum Fetuin B levels in mice fed chow or HFD for 12 and 18 weeks were analysed by western blot and normalized to Coomassie blue staining ( g , h ) (n = 4–6). The protein levels of Fetuin B in the liver of mice fed chow or HFD for 12 and 18 weeks were analysed by western blot and normalized to the loading control (β-actin) ( i , j ) (n = 5–6). The mRNA levels of Fetuin B in the livers of mice fed chow or a HFD for 12 weeks were analysed by qRT–PCR (k) (n = 5–6). Data are presented as mean ± SEM of all mice in each group. A nonparametric test (Mann–Whitney test) was applied. Significance is presented as * p < 0.05 compared with the chow group, ** p < 0.01 compared with the chow group, *** p < 0.001 compared with the chow group, **** p < 0.0001 compared with the chow group. Original blots are presented in the Supplementary Information file.

Article Snippet: The levels of human serum leptin (#ab179884), Fetuin B (#ab240684) and mouse serum leptin (#ab100718) were determined by ELISA (Abcam, Shanghai, China) according to the manufacturers’ instructions.

Techniques: Expressing, Western Blot, Staining, Quantitative RT-PCR, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Hepatokine Fetuin B expression is regulated by leptin-STAT3 signalling and associated with leptin in obesity

doi: 10.1038/s41598-022-17000-w

Figure Lengend Snippet: Effect of leptin and palmitic acid on Fetuin B expression in hepatocytes. The Fetuin B protein and mRNA levels were increased by treatment with leptin for 16 h in primary murine hepatocytes ( a , b ). The Fetuin B protein and mRNA levels were increased by treatment with leptin for 12 h in AML12 cells ( c , d ). Fetuin B protein and mRNA levels increased upon palmitic acid stimulation for 16 h in primary murine hepatocytes ( e , f ). The Fetuin B protein and mRNA levels were increased by palmitic acid stimulation for 12 h in AML12 cells. ( g , h ). The mRNA level of LepRb was analysed by qRT–PCR after palmitic acid stimulation in primary murine hepatocytes for 16 h ( i ) and AML12 cells for 12 h ( j ). qPCR data are presented as the mean ± SEM of two or three independent experiments. Western blot images are representative of two or more independent experiments. Quantitation of the representative western blot is presented as the mean ± SEM of triplicate measurements of the grey intensity ratio of Fetuin B/loading control. Original blots for independent replications for each experiment (n = 2–4) are presented in the Supplementary Information file. One-way analysis of variance (ANOVA) followed by Tukey’s multiple comparison test was used for experiments with three or more groups. t test was used to compare two groups. * p < 0.05 compared with the control group; ** p < 0.01 compared with the control group; *** p < 0.001 compared with the control group, **** p < 0.0001 compared with the control group.

Article Snippet: The levels of human serum leptin (#ab179884), Fetuin B (#ab240684) and mouse serum leptin (#ab100718) were determined by ELISA (Abcam, Shanghai, China) according to the manufacturers’ instructions.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Quantitation Assay

Journal: Scientific Reports

Article Title: Hepatokine Fetuin B expression is regulated by leptin-STAT3 signalling and associated with leptin in obesity

doi: 10.1038/s41598-022-17000-w

Figure Lengend Snippet: STAT3 phosphorylation mediates the transcription of FetuB in AML12 cells. STAT3 phosphorylation and Fetuin B protein expression levels in AML12 cells treated with leptin (50 ng/mL) for up to 6 h were analyzed by western blot ( a ). A Dual-Luciferase Reporter Assay was applied to determine FetuB promoter activity in AML12 cells by co-transfecting 100 ng luciferase reporter vector, 50 ng pRL-SV40-N and 100 ng pCMV-mSTAT3 or pCMV-HA. Firefly and Renilla luciferase activities were measured 36 h after transfection ( b ). ChIP assays were conducted to detect the binding of p-STAT3 to the FetuB promoter in AML12 cells treated with leptin treatment (50 ng/mL) for 12 h. PCR products were visualized by agarose gel electrophoresis and the efficiency of ChIP was calculated ( c ). STAT3 phosphorylation and Fetuin B levels in AML12 cells treated with or without leptin and Stattic treatments were analyzed by western blot ( d ). Data are presented as the mean ± SEM of three independent luciferase reporter assays and ChIP experiments. Quantitation of the representative western blot is presented as the mean ± SEM of triplicate measurements of grey intensities of targets. Original blots for independent replications for each experiment (n = 3) are presented in the Supplementary Information file. One-way analysis of variance (ANOVA) followed by Tukey’s multiple comparison test was used for experiments with three or more groups. t test was used to compare two groups. * p < 0.05 compared with the control group; ** p < 0.01 compared with the control group; *** p < 0.001 compared with the control group, **** p < 0.0001 compared with the control group.

Article Snippet: The levels of human serum leptin (#ab179884), Fetuin B (#ab240684) and mouse serum leptin (#ab100718) were determined by ELISA (Abcam, Shanghai, China) according to the manufacturers’ instructions.

Techniques: Expressing, Western Blot, Luciferase, Reporter Assay, Activity Assay, Plasmid Preparation, Transfection, Binding Assay, Agarose Gel Electrophoresis, Quantitation Assay

Journal: Scientific Reports

Article Title: Hepatokine Fetuin B expression is regulated by leptin-STAT3 signalling and associated with leptin in obesity

doi: 10.1038/s41598-022-17000-w

Figure Lengend Snippet: Leptin induces insulin resistance via Fetuin B. Mediation effects of serum Fetuin B on the association between serum leptin and HOMA-IR in obesity (n = 215). Sobel test was applied. Significance is presented as * p < 0.05, *** p < 0.001, and p < 0.05 for coefficients different from 0.

Article Snippet: The levels of human serum leptin (#ab179884), Fetuin B (#ab240684) and mouse serum leptin (#ab100718) were determined by ELISA (Abcam, Shanghai, China) according to the manufacturers’ instructions.

Techniques:

Journal: Scientific Reports

Article Title: Hepatokine Fetuin B expression is regulated by leptin-STAT3 signalling and associated with leptin in obesity

doi: 10.1038/s41598-022-17000-w

Figure Lengend Snippet: Demographic and clinical characteristics of subjects according to the tertiles of serum Fetuin B.

Article Snippet: The levels of human serum leptin (#ab179884), Fetuin B (#ab240684) and mouse serum leptin (#ab100718) were determined by ELISA (Abcam, Shanghai, China) according to the manufacturers’ instructions.

Techniques: