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: 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: Biomaterials

Article Title: Amino acid-based compound activates atypical PKC and leptin receptor pathways to improve glycemia and anxiety like behavior in diabetic mice.

doi: 10.1016/j.biomaterials.2020.119839

Figure Lengend Snippet: Fig. 4. AAC2 induced glucose uptake by LepR-dependent mechanism in vivo. (a) GTT in Leprdb mice treated without (Veh, open circles, n = 6) or with AAC2 (black circles, n = 6). Student's independent t-test for each time point. P = 0.006 at 30 min post glucose injection. (b,c) Food intake, P = 0.817, ns. (b) and weight gain P = 0.576, ns. (c) in Leprdb mice treated without (Veh, open bars, n = 6) or with AAC2 (black bar, n = 6). (d) Insulin levels in plasma of same Leprdb mice were measured by ELISA. P = 0.416, ns. (e) GTT in Lepob treated without (Veh, open circles, n = 5) or with AAC2 (black circles, n = 5). Student's independent t-test for each time point. P = 0.001 and P = 0.045 at 5 min and 30 min post glucose injection, respectively. (f–h) Food intake, P = 0.133, ns. (f), weight gain (g), and insulin levels (h) in Lepob mice treated without (Veh, open bar, n = 5) or with AAC2 (black bar, n = 5) P = 0.17, ns. Student's independent t-test.

Article Snippet: Interaction of either AAC2 (0.1 μM in PBS) or mouse recombinant leptin protein (mLep; 1.6 fM in PBS; Crystal Chem, Elk Grove Village, IL, 90030-B) and with mouse recombinant leptin receptor (LepR) protein (1.6 pM in PBS; R&D systems, 497-LR/CF) were investigated by applying an alternating current on quartz via the piezoelectric effect using QCM-D [52,53].

Techniques: In Vivo, Injection, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Degenerative Neurological and Neuromuscular Disease

Article Title: Neuroprotective Effects of Leptin on the APP/PS1 Alzheimer’s Disease Mouse Model: Role of Microglial and Neuroinflammation

doi: 10.2147/dnnd.s427781

Figure Lengend Snippet: Figure 1 Influence of leptin on the levels of Aβ1-40 (Left) and Aβ1-42 (Right) in the adult + leptin group, adult + saline group, aged + leptin group and aged + saline group.

Article Snippet: Recombinant mouse leptin (Elabscience.

Techniques: Saline

Journal: Degenerative Neurological and Neuromuscular Disease

Article Title: Neuroprotective Effects of Leptin on the APP/PS1 Alzheimer’s Disease Mouse Model: Role of Microglial and Neuroinflammation

doi: 10.2147/dnnd.s427781

Figure Lengend Snippet: Figure 2 Western blotting analysis of synaptophysin expression in the adult + leptin group, adult + saline group, aged + leptin group and aged + saline group.

Article Snippet: Recombinant mouse leptin (Elabscience.

Techniques: Western Blot, Expressing, Saline

Journal: Degenerative Neurological and Neuromuscular Disease

Article Title: Neuroprotective Effects of Leptin on the APP/PS1 Alzheimer’s Disease Mouse Model: Role of Microglial and Neuroinflammation

doi: 10.2147/dnnd.s427781

Figure Lengend Snippet: Figure 3 Semi-quantitative analysis of synaptophysin expression in the adult + leptin group, adult + saline group, aged + leptin group and aged + saline group.

Article Snippet: Recombinant mouse leptin (Elabscience.

Techniques: Expressing, Saline

Journal: Degenerative Neurological and Neuromuscular Disease

Article Title: Neuroprotective Effects of Leptin on the APP/PS1 Alzheimer’s Disease Mouse Model: Role of Microglial and Neuroinflammation

doi: 10.2147/dnnd.s427781

Figure Lengend Snippet: Figure 4 Leptin increased microglial cell number in the hippocampus of adult mice, but not aged mice. (A–D) Representative images of Iba-1+ cells, in the hippocampus sections of the mice. (E–G) Quantitative analysis of Iba-1+ stained cells.

Article Snippet: Recombinant mouse leptin (Elabscience.

Techniques: Staining

Journal: Degenerative Neurological and Neuromuscular Disease

Article Title: Neuroprotective Effects of Leptin on the APP/PS1 Alzheimer’s Disease Mouse Model: Role of Microglial and Neuroinflammation

doi: 10.2147/dnnd.s427781

Figure Lengend Snippet: Figure 5 Influence of leptin on the levels of IL-1β (Left), IL-6 (Right) in the adult + leptin group, adult + saline group, aged + leptin group and aged + saline group.

Article Snippet: Recombinant mouse leptin (Elabscience.

Techniques: Saline

Journal: Degenerative Neurological and Neuromuscular Disease

Article Title: Neuroprotective Effects of Leptin on the APP/PS1 Alzheimer’s Disease Mouse Model: Role of Microglial and Neuroinflammation

doi: 10.2147/dnnd.s427781

Figure Lengend Snippet: Figure 6 Western blotting analysis of leptin receptor expression in the adult + leptin group, adult + saline group, aged + leptin group and aged + saline group.

Article Snippet: Recombinant mouse leptin (Elabscience.

Techniques: Western Blot, Expressing, Saline

Journal: Degenerative Neurological and Neuromuscular Disease

Article Title: Neuroprotective Effects of Leptin on the APP/PS1 Alzheimer’s Disease Mouse Model: Role of Microglial and Neuroinflammation

doi: 10.2147/dnnd.s427781

Figure Lengend Snippet: Figure 7 Semi-quantitative analysis of leptin receptor expression in the adult + leptin group, adult + saline group, aged + leptin group and aged + saline group.

Article Snippet: Recombinant mouse leptin (Elabscience.

Techniques: Expressing, Saline

Journal: Nature communications

Article Title: Pharmacological and physiological activation of TGR5 in the NTS lowers food intake by enhancing leptin-STAT3 signaling.

doi: 10.1038/s41467-025-60331-1

Figure Lengend Snippet: Fig. 2 | Activation of TGR5 by CCDC enhances leptin-LepR-STAT3 signaling in the NTS to lower food intake. a Cumulative food intake of NTS CMC (n = 15; data replotted from Fig. 1b, h), leptin (n = 10; data replotted from Fig. 1h), CCDC (n = 10; data replotted from Fig. 1b) or CCDC+leptin (n = 11) HF rats 24 h after food was given back. b Representative western blot images of p-STAT3, t-STAT3, and GAPDH protein levels in NTS CCDC 5 min acute infusion, followed by a 3 h wait, then NTS CMC or leptin 5 min acute infusion, and a subsequent 30 min waiting time in HF rats. c Representative images (scale bar: 100 μm) of immunohistological staining for p-STAT3 with DAPI (top) and cFOS (bottom) in NTS CCDC 5 min acute infusion, followed by a 3 h wait, then NTS CMC or leptin 5 min acute infusion, and a sub- sequent 30 min waiting time in HF rats. d Relative NTS Tgr5 mRNA expression of NTS shMM (n = 6) or shTgr5 (n = 6) rats. e Relative NTS PKA activity of NTS CMC shMM (n = 6), CCDC shMM (n = 6), CMC shTgr5 (n = 6), or CCDC shTgr5 (n = 6) HF rats, measured 3 h after 5 min acute infusions. f Cumulative food intake of NTS CMC shMM (n = 5), CCDC+leptin shMM (n = 10), CMC shTgr5 (n = 5), or CCDC+leptin

Article Snippet: Leptin (497-LR, R&D Systems) was dissolved firstly in 0.9% saline and thenwith/withoutCCDC (final CCDCconcentration 100μM) in 1% CMC to the concentration of 153μM (equivalent to 0.5μg/site) as this amount of leptin infused into the hypothalamus or NTS lowers food intake23,64,65.

Techniques: Activation Assay, Western Blot, Staining, Expressing, Activity Assay

Journal: Nature communications

Article Title: Pharmacological and physiological activation of TGR5 in the NTS lowers food intake by enhancing leptin-STAT3 signaling.

doi: 10.1038/s41467-025-60331-1

Figure Lengend Snippet: Fig. 3 | Activation of TGR5 decreases inflammation and leptin signaling sup- pressor in the NTS. Relative NTS Cd68 (a), Il6 (b), Tnfa (c), LepRb (d), Ptp1b (e), and Socs3 (f) mRNA expression of NTS CMC (n = 8) or CCDC (n = 6) chow rats and NTS CMC (n = 8 for a–d; n = 7 for e, f) or CCDC (n = 8 for a–d; n = 7 for e, f) HF rats, measured 24 h after food was given back following 5 min acute infusion. g Representative images (scale bar: 100 μm; 25 μm for the digital zoomed-in ima- ges) showing FISH for Tgr5, Aif1, and DAPI or LepR, Aif1, and DAPI in the NTS of HF rats 30 min after CMC 5 min acute infusion. Digital zooms of the boxed regions are shown to the left. Arrowheads indicate cells co-expressing Tgr5 or LepR and Aif1. h Left: Proportion of Aif1+ microglia in the NTS that are Tgr5+ or Tgr5- and LepR+ or LepR- (pie chart). Right: Percentage of Aif1+ microglia expressing Tgr5or LepR in the NTS (n = 3 per group). i Representative images (scale bar: 100 μm; 25 μm for the digital zoomed-in images) showing FISH for Tgr5, Rbfox3, and DAPI or LepR, Rbfox3, and DAPI in the NTS of HF rats 30 min after CMC 5 min acute infusion.

Article Snippet: Leptin (497-LR, R&D Systems) was dissolved firstly in 0.9% saline and thenwith/withoutCCDC (final CCDCconcentration 100μM) in 1% CMC to the concentration of 153μM (equivalent to 0.5μg/site) as this amount of leptin infused into the hypothalamus or NTS lowers food intake23,64,65.

Techniques: Activation Assay, Expressing

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Pluronic modified leptin with increased systemic circulation, brain uptake and efficacy for treatment of obesity.

doi: 10.1016/j.jconrel.2014.05.044

Figure Lengend Snippet: Fig. 3. Purification and characterization of leptin–P85 conjugates. (A) Lep(ss)–P85 conjugate contained unmodified leptin and leptin modified with different numbers of P85 chains. SEC elution profile in TSKgel G2000SW column showed separation of leptin–P85 conjugates from unmodified leptin. (B) SDS-PAGE and (C) MALDI-TOF spectra further characterized the col- lected fractions at 8.8 min and 9.5 min as leptin conjugates modified with multiple P85 chains (Lep(ss)–P85(H)) and single P85 chain (Lep(ss)–P85(L)), respectively.

Article Snippet: Mouse recombinant leptin (Lep) and a chimera leptin receptor (ObRFc) were purchased from R&D Systems (Minneapolis, MN).

Techniques: SDS Page

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Pluronic modified leptin with increased systemic circulation, brain uptake and efficacy for treatment of obesity.

doi: 10.1016/j.jconrel.2014.05.044

Figure Lengend Snippet: Fig. 5. Disulfide bond stability in Lep(ss)–P85 conjugate upon its exposure to serum. Western blot analysis shows that leptin or Lep(ss)–P85 remained stable after incubating with serum for up to 24 hr.

Article Snippet: Mouse recombinant leptin (Lep) and a chimera leptin receptor (ObRFc) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Western Blot

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Pluronic modified leptin with increased systemic circulation, brain uptake and efficacy for treatment of obesity.

doi: 10.1016/j.jconrel.2014.05.044

Figure Lengend Snippet: Fig. 7. Serum clearance of leptin–pluronic conjugates during shorter (A and B) and longer time periods of study (C and D). The two lines in panels A and B were significantly different (p b 0.05). Both 125I-Lep(ss)–P85(L) and 125I-Lep(ss)–P85(H) showed significantly longer circulation time than that of co-injected 131I-leptin. The serum disappearance (T1/2) was 40.75 min for 125I-Lep(ss)–P85(L) (r = 0.75, p b 0.001; n = 1 ~ 2 mice/time point), 75.80 min for 125I-Lep(ss)–P85(H) (r = 0.73, p b 0.0005; n = 1 ~ 2 mice/time point) and 11.98 min for 131I-Lep (r = 0.64, p p b 0.005; n = 1 ~ 2 mice/time point). The vascular volume distribution (Vi), as shown by the y intercept was not significantly different. Panels C and D further showed that initial serum clearance was a linear distribution for both of leptin analogs followed by a plateau phase.

Article Snippet: Mouse recombinant leptin (Lep) and a chimera leptin receptor (ObRFc) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Injection

Journal: Scientific Reports

Article Title: The antidepressant fluoxetine acts on energy balance and leptin sensitivity via BDNF

doi: 10.1038/s41598-018-19886-x

Figure Lengend Snippet: FLX treated mice show enhanced response to acute leptin administration. (A) 14-h cumulative food intake in H 2 O-CFD and FLX-CFD mice at baseline and in response to i.p. leptin (3 mg/kg) injection. All data are expressed as % of the mean of the baseline of H 2 O-CFD. RM 2-way ANOVA (treatment x stimulus, F(1,45) = 7.93, P = 0.0072; matching, F(45,45) = 2.77, P = 0.0004; n = 22–25) followed by Bonferroni posthoc test, §§§ P< 0.001 versus baseline. (B) Left representative immunofluorescence showing expression of phosphorylated-STAT3 45′ minutes after an injection of saline (sal) or leptin (lep, 3 mg/kg) in the ARC of H 2 O-CFD (CNT) and FLX-CFD (FLX). Scale bar is 100 μm. Right , signal for pSTAT3 normalized to that of STAT3 was acquired for H 2 O- and FLX-CFD mice. All data are expressed as % of the mean of the H 2 O-CFD injected with saline. 2-way ANOVA (stimulus effect, F(1,34) = 77.18, P < 0.0001; treatment effect, F(1,34) = 14.06, P = 0.0007; n = 11–8, 10–9) followed by Bonferroni posthoc test, **P < 0.01 versus H 2 O-CFD, §§§ P< 0.001 versus saline. (C) OBRb mRNA value was determined by quantitative reverse transcription–PCR and standardized to TATA Binding Protein ( TBP ) in the hypothalamus of H 2 O-CFD and FLX-CFD mice. Student’s t-test (t = 3.212 df = 14, **P = 0.0063; n = 8). Data are presented as mean ± s.e.m.

Article Snippet: Food intake was measured 14 and 24 hours later in basal conditions, when mice were i.p. injected with recombinant mouse leptin (3 mg/Kg body weight, AstraZeneca).

Techniques: Injection, Immunofluorescence, Expressing, Binding Assay