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: 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