Fig 1: Impact of rCGH supplementation on metabolic parameters in ob/ob mice and OVX mice.(A) Quantification of GPHB5 expression in the skeletal muscle of mice with diet-induced obesity compared with control mice (n = 6 per group). (B–E) Energy expenditure (B), core body temperature (C), food intake (D), and body weight change (E) of ob/ob mice treated with vehicle (Veh) or 5 mg/kg rCGH (n = 4 per group). (F) Fat mass and lean mass of ob/ob mice treated with vehicle or 5 mg/kg rCGH (n = 4 per group). (G–J) Energy expenditure (G), food intake (H), body weight change (I), and fat mass (J) of OVX mice treated with vehicle or 5 mg/kg rCGH (n = 4–6 per group). All data represent mean ± SEM; significant differences were determined by unpaired 2-tailed Student’s t test (A, C, D, and F); ANCOVA with energy expenditure as a dependent variable and body weight as covariate (B and G); 1-way ANOVA with Dunnett’s multiple comparisons test (H and J); and 2-way ANOVA with Šídák’s multiple comparisons test (E and I). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. HFD, high-fat diet.
Fig 2: Mice lacking GPHB5 had decreased body temperature, browning, and fat metabolism.(A) Core body temperature of mice after cold exposure (4°C, 24 hours; n = 7 per group). (B and C) qPCR analysis of browning and lipolysis-associated genes, n = 6–7 per group (B) and Western blot analysis of UCP1, n = 7 for WT and n = 6 for GPHB5–/– mice (C) in iWAT after cold exposure (4°C, 24 hours). Quantification of UCP1 protein levels also is shown. (D) Representative images of H&E-stained iWAT and UCP1-specific IHC of iWAT from mice after cold exposure (4°C, 24 hours; n = 7 per group). Scale bars: 100 μm. (E) Metabolite heatmaps of mice iWAT (n = 8 per group) at room temperature (22°C). Rows reflected normalized (z-score) metabolite concentrations. (F and G) qPCR analysis of lipolysis and GLUTs associated genes in iWAT (F) and eWAT (G) of mice (n = 7 per group) at room temperature (22°C). All data represent mean ± SEM; significant differences between treatments were determined by unpaired 2-tailed Student’s t test (A and C) or Mann-Whitney U test (B, F, and G). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
Fig 3: Lack of GPHB5 led to glucose intolerance, insulin resistance, and hepatic lipid accumulation.Glucose tolerance test (GTT) (A) and insulin tolerance test (ITT) results (B) of mice (n = 5 for WT and n = 4–5 for GPHB5–/– mice). (C) Representative images of H&E-stained liver sections from WT and GPHB5–/– mice. Scale bars: 100 μm. (D) TG levels in liver of mice (n = 7 per group). (E) qPCR analysis of FA uptake– and TG synthesis–associated genes in liver of mice (n = 7 per group). All data represent mean ± SEM; significant differences between treatments were determined by 2-way ANOVA with Šídák’s multiple comparisons test (A and B); unpaired 2-tailed Student’s t test (D); and Mann-Whitney U test (E). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
Fig 4: Lack of GPHB5 expression led to obesity in mice.(A) Pictures of male WT mice (left) and GPHB5–/– mice (right) at 35 weeks of age. (B) Body weight curve for male mice (n = 18 for WT and n = 16 for GPHB5–/– mice). The body weight of mice was tracked weekly from the age of 6 weeks to 31 weeks. (C) Energy expenditure in mice (n = 8 per group) was measured by indirect calorimetry and analyzed by ANCOVA. (D) Body temperature of mice (n = 8 per group) at room temperature (22°C). (E) Body fat mass and lean mass of mice were detected by EchoMRI (n = 6 per group). (F) Micro-CT images of WT mice (left) and GPHB5–/– mice (right). The yellow parts in the coronal plane image represents WAT. (G) BAT between scapula, visceral fat (vis. fat), and subcutaneous fat (sub. fat) of mice (n = 5 per group) by micro-CT. All data represent mean ± SEM; significant differences were determined by 2-way ANOVA with Šídák’s multiple comparisons test (B); ANCOVA with energy expenditure as a dependent variable and body weight as covariate (C); and Student’s 2-tailed, unpaired t test (D, E, and G). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
Fig 5: Testosterone regulated the production of GPHB5.The testosterone levels in serum (A) and GPHB5 mRNA levels in brain (B) of male mice (n = 12 per group). (C) GPHB5 mRNA levels in brain, pituitary, and testis of mice in which AR was inhibited by flutamide for 10 days and 21 days (n = 4–5 per group). Con, control. (D) GPHB5 mRNA levels in brain, pituitary, and testis of mice (n = 6 for WT and n = 8 for AR–/– mice). (E) GPHB5 mRNA levels in C2C12 cells treated with testosterone for 1 day. (F) Relative luciferase activities in the C2C12 cells transfected with the WT and mutation GPHB5 promoter luciferase reporter plasmid. Cells were treated with DHT (10 nM) or enzalutamide (Enza; 0.1 μM) or both. NC, negative control. All data represent mean ± SEM; significant differences were determined by unpaired 2-tailed Student’s t test (A, B, and D); 1-way ANOVA with Dunnett’s multiple comparisons test (C and E); and 2-way ANOVA with Tukey’s multiple comparisons test (F). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
Supplier Page from MyBioSource.com for Human Glycoprotein hormone beta-5 (GPHB5) ELISA Kit