Journal: Nature Communications

Article Title: Adipocyte OGT governs diet-induced hyperphagia and obesity

doi: 10.1038/s41467-018-07461-x

Figure Lengend Snippet: Adipocyte OGT drives diet-induced hyperphagia and obesity. a Genetic cross scheme. b Growth curves and c body composition of adipocyte-specific OGT knockout mice (FKO) and littermate control mice (Con) on chow ( n = 8 (Con, dashed line with empty square/stripped bar) or 11 (FKO, blue line/bar)) or HFD ( n = 11 (Con, dashed line with empty circle/empty bar) or 13 (FKO, red line/bar)). d Circulating levels of leptin and e wet tissue weights in male Con ( n = 6) and FKO ( n = 6) mice fed HFD for 9 weeks, or age-matched Con ( n = 8) and FKO ( n = 8) mice fed chow. BAT, interscapular brown adipose tissue. White adipose tissues include depots from perigonadal (pg), subcutaneous (sc), and retroperitoneal (rp) regions. f Energy intake of male mice on HFD ( n = 7 per genotype) or Chow ( n = 5 (Con) or n = 6 (FKO)) for 13 weeks. g Fecal energy content. h Daily energy intake. Male Con ( n = 10) and FKO ( n = 10) mice fed HFD for 6 weeks. i Energy expenditure of 5-week HFD-fed male Con ( n = 7) and FKO ( n = 9) mice. j Energy intake in a two-choice diet preference test. Male Con ( n = 9) and FKO ( n = 10) mice have been fed HFD for 7 or 16 weeks. k Growth curves of HFD-fed mice. Con (AL, ad libitum, n = 11), Con (PF, pair-fed to littermate FKO, n = 7), FKO (AL, n = 7). Data were presented as mean ± s.e.m. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, post hoc Sidak’s tests ( b (HFD Con vs. HFD FKO), i , k (Con PF vs. FKO AL)), Tukey’s tests ( c , d , f ), or two-tailed Student’s t-test ( e , g , h , j )

Article Snippet: Recombinant mouse leptin protein (5 mg/kg, R&D Systems cat# 498-OB) was reconstituted at 1 mg/mL in sterile 20 mM Tris-HCl, pH 8.0.

Techniques: Knock-Out, Control, Two Tailed Test

Journal: Nature Communications

Article Title: Adipocyte OGT governs diet-induced hyperphagia and obesity

doi: 10.1038/s41467-018-07461-x

Figure Lengend Snippet: Adipocyte OGT drives diet-induced hyperphagia through transactivation of lipid desaturation in adipose tissue. a Gene expression analysis of de novo lipid desaturation and synthesis in adipose tissue (Chow for 15 weeks, Con n = 8, FKO n = 8; age-matched HFD for 9 weeks, Con n = 5, FKO n = 6). b Protein levels of SCD and OGT in adipose tissue. c Western blotting analysis in adipose tissue from 9-week-HFD-fed FKO and Con mice (Con n = 5, FKO n = 6, two representative biological replicates were shown). d Growth curves and e energy intake of (m)HFD-fed Con and FKO mice ( n = 8–10 per group). mHFD denotes a mono-unsaturated fat-fortified high-fat diet that changes the saturated fat-rich soybean oil fraction into the mono-unsaturated fat-rich canola oil as illustrated in the diagram ( d ). f Contents of palmitate (C16:0) and palmitoleate (C16:1) DAG in adipose lipidome, and circulating levels of leptin ( g ), or of FGF21 ( h ) in mice fed HFD or mHFD for 15 weeks. Data were presented as mean ± s.e.m. n.s. not significant, * P < 0.05, ** P < 0.01, **** P < 0.0001, post hoc Sidak’s tests ( a , e – h ), or # P < 0.01 HFD Con vs. HFD FKO, two-tailed Student’s t-test ( a ). A dashed line in ( e – h ) indicates that data from HFD study and mHFD study are presented in a side-by-side manner, and inference may not be made between different diets

Article Snippet: Recombinant mouse leptin protein (5 mg/kg, R&D Systems cat# 498-OB) was reconstituted at 1 mg/mL in sterile 20 mM Tris-HCl, pH 8.0.

Techniques: Gene Expression, Western Blot, Two Tailed Test

Journal: PLoS Genetics

Article Title: Congenital lipodystrophy induces severe osteosclerosis

doi: 10.1371/journal.pgen.1008244

Figure Lengend Snippet: A) Transplanted fat depots 3 months after surgery. B) Fat depot weight 3 months after transplantation. C) Serum leptin and adiponectin of FF mice 3 months after fat depot transplantation. WT and non-transplanted FF mice serve as control. D) μCT analysis of trabecular bone volume and bone mineral density of distal femurs of FF mice 3 months after sham operation or transplantation of fat derived from WT or adipokine-deficient mice. Data are presented as mean ± SD. **p<0.01; *** p<0.001 as determined by ANOVA with Holm-Sidak's post hoc analysis for multiple comparisons test. D) comparison with FF Sham except where detailed.

Article Snippet: The primary antibody cocktail contained rat anti-mouse CD45-BUV395 (BD Horizon, clone 30-F11, final dilution factor 1:200), rat anti-mouse TER-119-APC (BioLegend, clone TER-119, 1:200), rat anti-mouse CD41-BV421 (BioLegend, clone MWReg30, 1:300), rat anti-mouse/human CD11b (BioLegend, clone M1/70, 1:400), and rat-anti mouse Leptin receptor (LepR)-biotin (R&D Systems, polyclonal, 1:50) in Brilliant Stain Buffer (BD Biosciences) containing 10 μg/mL FcBlock.

Techniques: Transplantation Assay, Control, Derivative Assay, Comparison

Journal: PLoS ONE

Article Title: Long-lived weight-reduced αMUPA mice show higher and longer maternal-dependent postnatal leptin surge

doi: 10.1371/journal.pone.0188658

Figure Lengend Snippet: Each time point represents the mean ± s.e.m. of 5 to 17 mice (see for details). Pups were sampled every four days, following four hours of chow deprivation (first experiment), and were weaned at day 24 (P24). (A) Postnatal circulating leptin level measured using ELISA kit (R&D Systems Inc.). Insert of A. Mean P4-P24 leptin level. Bars with different letters are significantly different ( P <0.05, based on two-ways ANOVA followed by a post-hoc test). (B) Leptin:FM ratio. *, P <0.05 by post hoc comparison following one-way ANOVA of all four mice groups.

Article Snippet: Leptin level in the serum, milk, and stomach content was measured using a mouse leptin ELISA kit (R&D Systems Inc.), as used previously for both strains [ , ].

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: PLoS ONE

Article Title: Long-lived weight-reduced αMUPA mice show higher and longer maternal-dependent postnatal leptin surge

doi: 10.1371/journal.pone.0188658

Figure Lengend Snippet: Multiple regression of leptin on strain, gender, age, fat mass, and fat-free mass.

Article Snippet: Leptin level in the serum, milk, and stomach content was measured using a mouse leptin ELISA kit (R&D Systems Inc.), as used previously for both strains [ , ].

Techniques:

Journal: PLoS ONE

Article Title: Long-lived weight-reduced αMUPA mice show higher and longer maternal-dependent postnatal leptin surge

doi: 10.1371/journal.pone.0188658

Figure Lengend Snippet: Multiple regression of leptin on age, fat mass, strain, and gender, carried out separately for each age group.

Article Snippet: Leptin level in the serum, milk, and stomach content was measured using a mouse leptin ELISA kit (R&D Systems Inc.), as used previously for both strains [ , ].

Techniques:

Journal: PLoS ONE

Article Title: Long-lived weight-reduced αMUPA mice show higher and longer maternal-dependent postnatal leptin surge

doi: 10.1371/journal.pone.0188658

Figure Lengend Snippet: Each time point represents the mean ± s.e.m. of 6 to 22 mice, females only. Pups were sampled every four days, following four hours of chow deprivation (first and second experiments) and dam deprivation (second experiment only). Leptin and corticosterone levels were measured using ELISA kits (R&D Systems Inc.). (A and B) Postnatal circulating leptin level of dam-deprived and non-deprived female WT (left) and αMUPA (right) mice. (C) Postnatal circulating leptin levels of dam-deprived female αMUPA and WT mice. (D) Postnatal body weight growth curves of dam-deprived and non-deprived female WT mice. (E) Postnatal body weight growth curves of dam-deprived and non-deprived female αMUPA mice. (F) Postnatal body weight growth curves of dam-deprived female αMUPA and WT mice. (G) Mean P4-P24 leptin level of dam-deprived and non-deprived female αMUPA and WT mice. (H) Corticosterone P12 level of dam-deprived and non-deprived female αMUPA and WT mice. Bars with different letters are significantly different ( P <0.05, based on two-ways ANOVA followed by a post-hoc test). *, P <0.05 by post hoc comparison following one-way ANOVA of all four mice groups.

Article Snippet: Leptin level in the serum, milk, and stomach content was measured using a mouse leptin ELISA kit (R&D Systems Inc.), as used previously for both strains [ , ].

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: PLoS ONE

Article Title: Long-lived weight-reduced αMUPA mice show higher and longer maternal-dependent postnatal leptin surge

doi: 10.1371/journal.pone.0188658

Figure Lengend Snippet: Bars represent the mean ± s.e.m. of 6 dams per strain (A) and their female pups (B), one female pup per dam, all sampled at P12. Dams' serum and milk samples (A) were obtained 4 hours after their separation from their pups. Milk secretion was encouraged by subcutaneous oxytocin injection (2 IU/kg) administered 15 min before sampling. Pups' serum and stomach content (B) were sampled 30 min after they were reunited with their dams. Leptin level was measured using ELISA kit (R&D Systems Inc.). *, P<0.05 between strains, by Mann-Whitney non-parametric test. #, P<0.05 within strains, by Wilcoxon non-parametric Test.

Article Snippet: Leptin level in the serum, milk, and stomach content was measured using a mouse leptin ELISA kit (R&D Systems Inc.), as used previously for both strains [ , ].

Techniques: Injection, Sampling, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: American journal of physiology. Gastrointestinal and liver physiology

Article Title: Leptin regulates gallbladder genes related to absorption and secretion.

doi: 10.1152/ajpgi.00389.2006

Figure Lengend Snippet: Fig. 1. Leptin administration decreased gallbladder volume, bile sodium concentration, and pH.

Article Snippet: For leptin replacement, mice received daily intraperitoneal injections of either recombinant murine leptin (R&D Systems, Minneapolis, MN) at a dose of 5 g/g body wt or saline as a control for 4 wk.

Techniques: Concentration Assay

Journal: PLoS ONE

Article Title: Chito-Oligosaccharide Inhibits the De-Methylation of a ‘CpG’ Island within the Leptin ( LEP ) Promoter during Adipogenesis of 3T3-L1 Cells

doi: 10.1371/journal.pone.0060011

Figure Lengend Snippet: Mouse 33-L1 pre-adipocytes were induced to differentiate and cells were harvested for RNA isolation and quantitative expression of LEP gene on days 0, 2, 4 and 6 post-induction of differentiation. Cell culture media was harvested for quantification of leptin production on days 2, 4 and 6 post-induction of differentiation. Day 0 and day 6 correspond to pre-adipocyte and mature adipocytes, respectively. Day 2 and day 4 correspond to a phase where the cells were actively involved in the differentiating process. Data represents mean ± standard error from three independent replicate experiments.

Article Snippet: Leptin was measured in the supernatant on day 0, 2, 4 and 6PID in absence or presence of COS. Leptin was quantified using a mouse leptin sandwich ELISA (R&D Systems Europe, Ltd. Abingdon, UK) according to the manufacturer’s instructions.

Techniques: Isolation, Expressing, Cell Culture

Journal: PLoS ONE

Article Title: Chito-Oligosaccharide Inhibits the De-Methylation of a ‘CpG’ Island within the Leptin ( LEP ) Promoter during Adipogenesis of 3T3-L1 Cells

doi: 10.1371/journal.pone.0060011

Figure Lengend Snippet: Mouse 33-L1 pre-adipocytes were induced to differentiate in the absence or presence of chitooligosaccharide and cell culture media was harvested for quantification of leptin production on days 2–4 post-induction of differentiation (A) and days 4–6 post-induction of differentiation (B) Data represents mean ± standard error from three independent replicate experiments. Note the differences in the scale of Y-axis representing the leptin concentration (pg/ml) in the media for different figures.

Article Snippet: Leptin was measured in the supernatant on day 0, 2, 4 and 6PID in absence or presence of COS. Leptin was quantified using a mouse leptin sandwich ELISA (R&D Systems Europe, Ltd. Abingdon, UK) according to the manufacturer’s instructions.

Techniques: Cell Culture, Concentration Assay