Journal: Nature communications

Article Title: A mouse model for a partially inactive obesity-associated human MC3R variant.

doi: 10.1038/ncomms10522

Figure Lengend Snippet: Figure 8 | Maintained adipose tissue function in MC3RhDM/hDM mice. Gonadal fat was isolated from fasted female mice fed a high-fat diet for (a) quantitative real-time PCR (MC3RhWT/hWT n ¼ 5; MC3RhDM/hDM n ¼ 8/group), (b–d) western blotting analysis (MC3RhWT/hWT n ¼ 5; MC3RhDM/hDM n ¼ 4), and (e–l) FACS analysis (MC3RhWT/hWT n ¼ 4; MC3RhDM/hDM n ¼ 5) in MC3RhWT/hWT (open bars) and MC3RhDM/hDM (closed bars) mice. (a) The expression levels of genes related to macrophage infiltration, inflammation and adipose tissue metabolism were normalized for b-actin expression. Western protein expression results for (b) peroxisome proliferator-activated receptor gamma (PPARg), (c) adiponectin and (d) phosphospecific 50-adenosine monophosphate- activated protein kinase (p-AMPK), divided by glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression and normalized relative to average for MC3RhWT/hWT. The stromal vascular fraction was isolated from gonadal fat (B2 g) from MC3RhWT/hWTand MC3RhDM/hDMmice. For detecting macrophages by flow cytometry, a gating strategy was used to enrich samples from MC3RhWT/hWTand MC3RhDM/hDMmice for adipose tissue macrophages by selecting cells in the gate 1 area (e,g). The gate 1 area was stained with a F4/80 antibody (f and h) and the dot plots depict forward scatter (FSC) and side scatter (SSC) (left). For detecting neutrophils, whole cells (non-gated) were stained with CD11b and Gr-1 (i,j). Bar graphs show the average values±s.e.m. for percentage of cells in the stromal vascular fraction that were (k) macrophages and (l) neutrophils. *Po0.05 MC3RhDM/hDMversus MC3RhWT/hWT. (m,n) Haematoxylin and eosin stained sections showed no apparent differences in adipocyte morphology between groups. Groups were compared by Student’s t-tests (two-tailed) (a–d and k–i). Scale bar, 100 mm. Data are represented as mean±s.e.m. for a-d, k, and l. FACS, fluorescence-activated cell sorting

Article Snippet: Other metabolites or hormones were measured using the indicated kits; Insulin (SRI-13K, Linco Research, St Charles, MO), corticosterone (ADI-900-097; Enzo Life science, Farmingdale, NY), IGF-1 (22-IGF-R21, ALPCO, Salem, NH), Adiponectin (DRP 300 for human plasma samples obtained by venipuncture and MRP300 for mouse serum, R&D SYSTEMS, Minneapolis, MN), Leptin (MOB00, R&D SYSTEMS), triglycerides (337-B; Sigma), and non-esterified fatty acids (13831175; Roche Molecular Biochemicals, Indianapolis, IN).

Techniques: Isolation, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Cytometry, Staining, Two Tailed Test, FACS

Journal: Journal of Molecular Neuroscience

Article Title: Unraveling Molecular Mechanisms of THAP1 Missense Mutations in DYT6 Dystonia

doi: 10.1007/s12031-020-01490-2

Figure Lengend Snippet: THAP1 molecular weight and subcellular distribution of THAP1. a Western blot showing a 30-kDa band and a 25-kDa band in HEK cells overexpressing wild-type THAP1 or F21L mutant THAP1 using the Proteintech antibody (12584-1-AP). A small band (around 27 kDa) was observed in HEK cells overexpressing Q154fs180X mutant THAP1. Mass spectrometry analysis detected two THAP1 peptides in the 30-kDa band (peptide 1, IHQLEQQVEK; peptide 2, LKEVVHFQK) but not in the 25-kDa band. b Wild-type and missense mutant (F81L and L180S) THAP1 protein distributed mainly in the nuclear fraction. Lamin A/C was used as marker of nuclear protein

Article Snippet: Fig. 1 THAP1 molecular weight and subcellular distribution of THAP1. a Western blot showing a 30-kDa band and a 25-kDa band in HEK cells overexpressing wild-type THAP1 or F21L mutant THAP1 using the Proteintech antibody (12584-1-AP).

Techniques: Molecular Weight, Western Blot, Mutagenesis, Mass Spectrometry, Marker

Journal: World journal of gastroenterology

Article Title: Role of gut microbiota in Crohn's disease pathogenesis: Insights from fecal microbiota transplantation in mouse model.

doi: 10.3748/wjg.v30.i31.3689

Figure Lengend Snippet: Figure 1 Histopathological alterations in mesenteric adipose and intestinal tissues of patients with Crohn’s disease. A: Mesenteric adipose tissues from Crohn’s disease (CD) patients and non-CD subjects stained with hematoxylin-eosin to assess histopathological alterations (× 150); B: Real-time reverse transcriptase-polymerase chain reaction assays of the levels of expression of interleukin-6 (IL-6), IL-1β, tumour necrosis factor alpha, MCP-1, leptin, and adiponectin mRNAs; C: Intestinal tissues from CD patients and non-CD subjects stained with Masson stain to evaluate fibrotic alterations (× 150); D: Immunohistochemical staining of alpha-smooth muscle actin in intestinal tissues from CD patients and non-CD subjects (× 150). Average optical density was quantified using ImageJ software (right panel). n = 12, bP < 0.01 or n = 3, bP < 0.01 by paired t-tests. HE: Hematoxylin and eosin; TNBS: 2,4,6-trinitrobenzene sulfonic acid; CD: Crohn’s disease; TNF-α: Tumour necrosis factor alpha; IL: Interleukin; α-SMA: Alpha-smooth muscle actin; AOD: Average optical density.

Article Snippet: A 100 μL aliquot of serum was added to each well of an ELISA plate included in ELISA kits measuring serum concentrations of endotoxin (CSB-E13066m, CUSABIO, Wuhan, Hubei Province, China), IL-6 (CSB-E04639m, CUSABIO), IL-1β (Ab197742, Abcam, Cambridge, United Kingdom), TNF-α (CSB-E04741m, CUSABIO), MCP-1 (Ab208979, Abcam), leptin (ab100718, Abcam), and adiponectin (CSB-E07272m, CUSABIO).

Techniques: Staining, Reverse Transcription, Polymerase Chain Reaction, Expressing, Immunohistochemical staining, Software

Journal: World journal of gastroenterology

Article Title: Role of gut microbiota in Crohn's disease pathogenesis: Insights from fecal microbiota transplantation in mouse model.

doi: 10.3748/wjg.v30.i31.3689

Figure Lengend Snippet: Figure 4 ELISA determinations of serum concentrations of cytokines and adipokines. A: Interleukin (IL)-6; B: IL-1β; C: Tumour necrosis factor alpha; D: MCP-1; E: Leptin; F: Adiponectin. n = 6, bP < 0.01 vs control; aP < 0.05, bP < 0.01 vs 2,4,6-trinitrobenzene sulfonic acid (TNBS) alone; bP < 0.01 vs TNBS + NC-fetal microbiota transplantation, by one-way ANOVA with post-hoc Tukey HSD test. TNBS: 2,4,6-trinitrobenzene sulfonic acid; CD: Crohn’s disease; NC: Normal control; FMT: Fetal microbiota transplantation.

Article Snippet: A 100 μL aliquot of serum was added to each well of an ELISA plate included in ELISA kits measuring serum concentrations of endotoxin (CSB-E13066m, CUSABIO, Wuhan, Hubei Province, China), IL-6 (CSB-E04639m, CUSABIO), IL-1β (Ab197742, Abcam, Cambridge, United Kingdom), TNF-α (CSB-E04741m, CUSABIO), MCP-1 (Ab208979, Abcam), leptin (ab100718, Abcam), and adiponectin (CSB-E07272m, CUSABIO).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Transplantation Assay

Journal: World journal of gastroenterology

Article Title: Role of gut microbiota in Crohn's disease pathogenesis: Insights from fecal microbiota transplantation in mouse model.

doi: 10.3748/wjg.v30.i31.3689

Figure Lengend Snippet: Figure 5 Effects of gut microbiota from Crohn’s disease patients on the histopathological characteristics of mesenteric adipose and intestinal tissues from mice with 2,4,6-trinitrobenzene sulfonic acid-induced Crohn’s disease. A: Mesenteric adipose tissues collected from mice were stained with hematoxylin-eosin to assess histopathological alterations (× 150); B and C: Real-time reverse transcriptase-polymerase chain reaction assays of interleukin-6 (IL-6), IL-1β, tumour necrosis factor alpha, MCP-1, leptin, adiponectin, LXR, and FXR mRNA levels in mouse tissues; D: Intestinal tissues from mice stained with Masson stain to evaluate fibrotic alterations (× 150); E: Immunoblotting assays of alpha-smooth muscle actin and vimentin protein levels in mouse intestinal tissues. n = 6 or 3, bP < 0.01 vs control; aP < 0.05, bP < 0.01 vs 2,4,6-trinitrobenzene sulfonic acid (TNBS) alone; bP < 0.01 vs TNBS + NC-fetal microbiota transplantation, by one-way ANOVA with post-hoc Tukey HSD test. TNBS: 2,4,6-trinitrobenzene sulfonic acid; CD: Crohn’s disease; NC: Normal control; FMT: Fetal microbiota transplantation; IFN-γ: Interferon-gamma; TNF-α: Tumour necrosis factor alpha; IL: Interleukin; HE: Hematoxylin and eosin; α-SMA: Alpha-smooth muscle actin.

Article Snippet: A 100 μL aliquot of serum was added to each well of an ELISA plate included in ELISA kits measuring serum concentrations of endotoxin (CSB-E13066m, CUSABIO, Wuhan, Hubei Province, China), IL-6 (CSB-E04639m, CUSABIO), IL-1β (Ab197742, Abcam, Cambridge, United Kingdom), TNF-α (CSB-E04741m, CUSABIO), MCP-1 (Ab208979, Abcam), leptin (ab100718, Abcam), and adiponectin (CSB-E07272m, CUSABIO).

Techniques: Staining, Reverse Transcription, Polymerase Chain Reaction, Western Blot, Control, Transplantation Assay

Journal: Diseases

Article Title: Assessing Anti-Adipogenic Effects of Mango Leaf Tea and Mangiferin within Cultured Adipocytes

doi: 10.3390/diseases11020070

Figure Lengend Snippet: Incubating cultured adipocytes with MLT augments the secretion of adiponectin and enhances glucose uptake. ( A , B ) Adipocytes were treated with vehicle, MGF, or MLT. ELISA was used to measure secretory adiponectin ( A ), and a fluorometric kit was used for measuring glucose uptake of fluorescently labeled 2-NBDG ( B ). ( A ) Four independent experiments were conducted using one biological replicate for each experiment. ( B ) Two independent experiments were performed, each with triplicate biological replicates used. ( A , B ) Bars are group means.

Article Snippet: We measured adiponectin in the centrifuged cell culture medium using a mouse adiponectin ELISA kit (AccuSignal TM ELISA Kit, KOA0366; Rockland, Limerick, PA, USA), following the manufacturer’s instructions.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Labeling

Journal: Particle and Fibre Toxicology

Article Title: Programming of mouse obesity by maternal exposure to concentrated ambient fine particles

doi: 10.1186/s12989-017-0201-9

Figure Lengend Snippet: Maternal exposure to CAP decreases adipose leptin expression. a the plasma leptin levels of adult offspring. n Male/FA = 6, n Male/CAP = 7, n Female/FA = 4, and n Female/CAP = 7. * p < 0.05, one way ANOVA. b the plasma adiponectin levels of adult offspring. n Male/FA = 6, n Male/CAP = 7, n Female/FA = 4, and n Female/CAP = 7. * p < 0.05, one way ANOVA. c the mRNA expression levels of offspring epididymal adipose tissues. n Male/FA = 6, n Male/CAP = 7, n Female/FA = 4, and n Female/CAP = 7. * p < 0.05, one way ANOVA. d the representative image of western blot analysis of leptin protein in epididymal adipose tissues. e the quantification of leptin protein in epididymal adipose tissues. n Male/FA = 6, n Male/CAP = 7, n Female/FA = 4, and n Female/CAP = 7. * p < 0.05, one way ANOVA. f the average methylation level of leptin promoter in adipocytes. n Male/FA = 6, n Male/CAP = 7, n Female/FA = 4, and n Female/CAP = 7. * p < 0.05, one way ANOVA

Article Snippet: Plasma insulin (Ultra Sensitive Mouse Insulin ELISA Kit, Crystal Chemical), leptin (RayBio Mouse Leptin ELISA Kit, RayBiotech), and adiponectin (Mouse Adiponectin ELISA Kit, Boster Biological Technology) levels were determined per manufacturer’s instruction.

Techniques: Expressing, Clinical Proteomics, Western Blot, Methylation