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(A) ANCOVA of energy expenditure (EE) with body weight as a covariate during the 12-hour dark cycle. (B) Average EE during the dark cycle normalized to body weight. (C) Respiratory Exchange Ratio (RER) over a 24-hour period. (D) Average RER during the dark cycle. (E) Spontaneous activity over a 24-hour period. (F) Average spontaneous activity during the dark cycle, calculated as laser beam breaks. (G) Circulating <t>FGF21</t> expression quantified from refed plasma using ELISA. (H) The mRNA expression of Fgf21 in the liver of mice. (A-F) n=11-12 mice per group, (G-H) n=3-5 mice per group. (A) Data for each individual mouse is plotted, simple linear regression (ANCOVA) was calculated to determine if the slopes or elevations are equal. (B, D, F-H) Statistics for the overall effects of diet, gonadectomy, and the interaction represent the p value from a two-way ANOVA. *p<0.05 from a Šidák’s post-test examining the effect of parameters identified as significant in the two-way ANOVA. Data are represented as mean ±SEM. Abbreviations: CTL (21% control protein diet), LP (7% low protein diet), Cast (castration), Fem (female), Ovx (ovariectomy), BW (body weight), EE (energy expenditure), RER (respiratory exchange ratio), FGF21 (fibroblast growth factor 21).
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(A) ANCOVA of energy expenditure (EE) with body weight as a covariate during the 12-hour dark cycle. (B) Average EE during the dark cycle normalized to body weight. (C) Respiratory Exchange Ratio (RER) over a 24-hour period. (D) Average RER during the dark cycle. (E) Spontaneous activity over a 24-hour period. (F) Average spontaneous activity during the dark cycle, calculated as laser beam breaks. (G) Circulating <t>FGF21</t> expression quantified from refed plasma using ELISA. (H) The mRNA expression of Fgf21 in the liver of mice. (A-F) n=11-12 mice per group, (G-H) n=3-5 mice per group. (A) Data for each individual mouse is plotted, simple linear regression (ANCOVA) was calculated to determine if the slopes or elevations are equal. (B, D, F-H) Statistics for the overall effects of diet, gonadectomy, and the interaction represent the p value from a two-way ANOVA. *p<0.05 from a Šidák’s post-test examining the effect of parameters identified as significant in the two-way ANOVA. Data are represented as mean ±SEM. Abbreviations: CTL (21% control protein diet), LP (7% low protein diet), Cast (castration), Fem (female), Ovx (ovariectomy), BW (body weight), EE (energy expenditure), RER (respiratory exchange ratio), FGF21 (fibroblast growth factor 21).
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(A) ANCOVA of energy expenditure (EE) with body weight as a covariate during the 12-hour dark cycle. (B) Average EE during the dark cycle normalized to body weight. (C) Respiratory Exchange Ratio (RER) over a 24-hour period. (D) Average RER during the dark cycle. (E) Spontaneous activity over a 24-hour period. (F) Average spontaneous activity during the dark cycle, calculated as laser beam breaks. (G) Circulating <t>FGF21</t> expression quantified from refed plasma using ELISA. (H) The mRNA expression of Fgf21 in the liver of mice. (A-F) n=11-12 mice per group, (G-H) n=3-5 mice per group. (A) Data for each individual mouse is plotted, simple linear regression (ANCOVA) was calculated to determine if the slopes or elevations are equal. (B, D, F-H) Statistics for the overall effects of diet, gonadectomy, and the interaction represent the p value from a two-way ANOVA. *p<0.05 from a Šidák’s post-test examining the effect of parameters identified as significant in the two-way ANOVA. Data are represented as mean ±SEM. Abbreviations: CTL (21% control protein diet), LP (7% low protein diet), Cast (castration), Fem (female), Ovx (ovariectomy), BW (body weight), EE (energy expenditure), RER (respiratory exchange ratio), FGF21 (fibroblast growth factor 21).
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(A) ANCOVA of energy expenditure (EE) with body weight as a covariate during the 12-hour dark cycle. (B) Average EE during the dark cycle normalized to body weight. (C) Respiratory Exchange Ratio (RER) over a 24-hour period. (D) Average RER during the dark cycle. (E) Spontaneous activity over a 24-hour period. (F) Average spontaneous activity during the dark cycle, calculated as laser beam breaks. (G) Circulating <t>FGF21</t> expression quantified from refed plasma using ELISA. (H) The mRNA expression of Fgf21 in the liver of mice. (A-F) n=11-12 mice per group, (G-H) n=3-5 mice per group. (A) Data for each individual mouse is plotted, simple linear regression (ANCOVA) was calculated to determine if the slopes or elevations are equal. (B, D, F-H) Statistics for the overall effects of diet, gonadectomy, and the interaction represent the p value from a two-way ANOVA. *p<0.05 from a Šidák’s post-test examining the effect of parameters identified as significant in the two-way ANOVA. Data are represented as mean ±SEM. Abbreviations: CTL (21% control protein diet), LP (7% low protein diet), Cast (castration), Fem (female), Ovx (ovariectomy), BW (body weight), EE (energy expenditure), RER (respiratory exchange ratio), FGF21 (fibroblast growth factor 21).
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recombinant mouse fgf21 protein  (Shanghai Aladdin Bio-Chem)
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(A) ANCOVA of energy expenditure (EE) with body weight as a covariate during the 12-hour dark cycle. (B) Average EE during the dark cycle normalized to body weight. (C) Respiratory Exchange Ratio (RER) over a 24-hour period. (D) Average RER during the dark cycle. (E) Spontaneous activity over a 24-hour period. (F) Average spontaneous activity during the dark cycle, calculated as laser beam breaks. (G) Circulating <t>FGF21</t> expression quantified from refed plasma using ELISA. (H) The mRNA expression of Fgf21 in the liver of mice. (A-F) n=11-12 mice per group, (G-H) n=3-5 mice per group. (A) Data for each individual mouse is plotted, simple linear regression (ANCOVA) was calculated to determine if the slopes or elevations are equal. (B, D, F-H) Statistics for the overall effects of diet, gonadectomy, and the interaction represent the p value from a two-way ANOVA. *p<0.05 from a Šidák’s post-test examining the effect of parameters identified as significant in the two-way ANOVA. Data are represented as mean ±SEM. Abbreviations: CTL (21% control protein diet), LP (7% low protein diet), Cast (castration), Fem (female), Ovx (ovariectomy), BW (body weight), EE (energy expenditure), RER (respiratory exchange ratio), FGF21 (fibroblast growth factor 21).
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serum fgf21  (R&D Systems)
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(A) ANCOVA of energy expenditure (EE) with body weight as a covariate during the 12-hour dark cycle. (B) Average EE during the dark cycle normalized to body weight. (C) Respiratory Exchange Ratio (RER) over a 24-hour period. (D) Average RER during the dark cycle. (E) Spontaneous activity over a 24-hour period. (F) Average spontaneous activity during the dark cycle, calculated as laser beam breaks. (G) Circulating <t>FGF21</t> expression quantified from refed plasma using ELISA. (H) The mRNA expression of Fgf21 in the liver of mice. (A-F) n=11-12 mice per group, (G-H) n=3-5 mice per group. (A) Data for each individual mouse is plotted, simple linear regression (ANCOVA) was calculated to determine if the slopes or elevations are equal. (B, D, F-H) Statistics for the overall effects of diet, gonadectomy, and the interaction represent the p value from a two-way ANOVA. *p<0.05 from a Šidák’s post-test examining the effect of parameters identified as significant in the two-way ANOVA. Data are represented as mean ±SEM. Abbreviations: CTL (21% control protein diet), LP (7% low protein diet), Cast (castration), Fem (female), Ovx (ovariectomy), BW (body weight), EE (energy expenditure), RER (respiratory exchange ratio), FGF21 (fibroblast growth factor 21).
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(A) ANCOVA of energy expenditure (EE) with body weight as a covariate during the 12-hour dark cycle. (B) Average EE during the dark cycle normalized to body weight. (C) Respiratory Exchange Ratio (RER) over a 24-hour period. (D) Average RER during the dark cycle. (E) Spontaneous activity over a 24-hour period. (F) Average spontaneous activity during the dark cycle, calculated as laser beam breaks. (G) Circulating FGF21 expression quantified from refed plasma using ELISA. (H) The mRNA expression of Fgf21 in the liver of mice. (A-F) n=11-12 mice per group, (G-H) n=3-5 mice per group. (A) Data for each individual mouse is plotted, simple linear regression (ANCOVA) was calculated to determine if the slopes or elevations are equal. (B, D, F-H) Statistics for the overall effects of diet, gonadectomy, and the interaction represent the p value from a two-way ANOVA. *p<0.05 from a Šidák’s post-test examining the effect of parameters identified as significant in the two-way ANOVA. Data are represented as mean ±SEM. Abbreviations: CTL (21% control protein diet), LP (7% low protein diet), Cast (castration), Fem (female), Ovx (ovariectomy), BW (body weight), EE (energy expenditure), RER (respiratory exchange ratio), FGF21 (fibroblast growth factor 21).

Journal: bioRxiv

Article Title: Female resistance to the metabolic benefits of protein restriction is reversed by ovariectomy in mice

doi: 10.64898/2026.03.31.715667

Figure Lengend Snippet: (A) ANCOVA of energy expenditure (EE) with body weight as a covariate during the 12-hour dark cycle. (B) Average EE during the dark cycle normalized to body weight. (C) Respiratory Exchange Ratio (RER) over a 24-hour period. (D) Average RER during the dark cycle. (E) Spontaneous activity over a 24-hour period. (F) Average spontaneous activity during the dark cycle, calculated as laser beam breaks. (G) Circulating FGF21 expression quantified from refed plasma using ELISA. (H) The mRNA expression of Fgf21 in the liver of mice. (A-F) n=11-12 mice per group, (G-H) n=3-5 mice per group. (A) Data for each individual mouse is plotted, simple linear regression (ANCOVA) was calculated to determine if the slopes or elevations are equal. (B, D, F-H) Statistics for the overall effects of diet, gonadectomy, and the interaction represent the p value from a two-way ANOVA. *p<0.05 from a Šidák’s post-test examining the effect of parameters identified as significant in the two-way ANOVA. Data are represented as mean ±SEM. Abbreviations: CTL (21% control protein diet), LP (7% low protein diet), Cast (castration), Fem (female), Ovx (ovariectomy), BW (body weight), EE (energy expenditure), RER (respiratory exchange ratio), FGF21 (fibroblast growth factor 21).

Article Snippet: Blood for circulating FGF21, testosterone, and estradiol levels was collected from submandibular bleeding immediately prior to euthanasia; plasma was assayed with a mouse/rat FGF21 quantikine ELISA kit (MF2100) from R&D Systems (Minneapolis, MN, USA), and testosterone (582701) and estradiol (501890) quantikine ELISA kits from Caymen Chemicals (Ann Arbor, MI, USA).

Techniques: Activity Assay, Expressing, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control