Journal: Stem Cell Research & Therapy

Article Title: Therapeutic potential of small extracellular vesicles derived from lipoma tissue in adipose tissue regeneration—an in vitro and in vivo study

doi: 10.1186/s13287-021-02291-z

Figure Lengend Snippet: Primers used for RT-PCR

Article Snippet: Immunochemical staining for C/EBP-α (Bioss, China, cat.bs1630R), PPARγ (Bioss, China, cat.bs4888R), Adiponectin (Bioss, China, cat.bs0471R), and CD31 (Sino Biological, China, cat.50408-T16) was performed to determine the extent of adipose tissue and blood vessel formation in new growth tissues at the 4th week.

Techniques:

Journal: Stem Cell Research & Therapy

Article Title: Therapeutic potential of small extracellular vesicles derived from lipoma tissue in adipose tissue regeneration—an in vitro and in vivo study

doi: 10.1186/s13287-021-02291-z

Figure Lengend Snippet: The biological responses of ADSCs to sEV-LT and sEV-AT. a The uptake of sEV-LT and sEV-AT by ADSCs. ADSCs were incubated with DiO-labeled extracellular vesicles (green) and stained with phallotoxins (red). Nuclei were stained with DAPI (blue). Scale bar = 50 μm. b Representative images from the scratch wound assay. Scale bar = 200 μm. c Quantitative analysis of cell migration in each group at 12 h and 24 h ( n = 3). d CCK-8 assay ( n = 3). e ADSCs cultured with sEV-LT and sEV-AT for 14 days, and the lipid droplets were stained with Oil Red O to determine the level of adipogenesis. ADSCs cultured with basal culture medium (negative control, NC) or adipogenic medium (ADIPO) were used as a negative and positive controls, respectively. Scale bar = 50 μm. f Quantification of the amount of Oil Red O ( n = 3). g The relative expressions of mRNA encoding C/EBPδ, PPARγ2, Adiponectin was measured by RT-PCR on day 7 after induction. Results are present as mean ± s.d. ( n = 3). * p < 0.05, ** p < 0.01 and *** p < 0.001

Article Snippet: Immunochemical staining for C/EBP-α (Bioss, China, cat.bs1630R), PPARγ (Bioss, China, cat.bs4888R), Adiponectin (Bioss, China, cat.bs0471R), and CD31 (Sino Biological, China, cat.50408-T16) was performed to determine the extent of adipose tissue and blood vessel formation in new growth tissues at the 4th week.

Techniques: Incubation, Labeling, Staining, Scratch Wound Assay Assay, Migration, CCK-8 Assay, Cell Culture, Negative Control, Reverse Transcription Polymerase Chain Reaction

Journal: Stem Cell Research & Therapy

Article Title: Therapeutic potential of small extracellular vesicles derived from lipoma tissue in adipose tissue regeneration—an in vitro and in vivo study

doi: 10.1186/s13287-021-02291-z

Figure Lengend Snippet: Immunohistochemistry of neotissue sections in the experimental and control groups at week 4 after transplantation. Representative photographs of C/EBPα, PPARγ, Adiponectin, and CD31 immunostaining in the Matrigel group, sEV-LT group, and sEV-AT group, respectively. The black arrows indicated the positive stained protein. Scale bar = 50 μm

Article Snippet: Immunochemical staining for C/EBP-α (Bioss, China, cat.bs1630R), PPARγ (Bioss, China, cat.bs4888R), Adiponectin (Bioss, China, cat.bs0471R), and CD31 (Sino Biological, China, cat.50408-T16) was performed to determine the extent of adipose tissue and blood vessel formation in new growth tissues at the 4th week.

Techniques: Immunohistochemistry, Transplantation Assay, Immunostaining, Staining

Journal: Journal of Diabetes Investigation

Article Title: Growth arrest‐specific 6 modulates adiponectin expression and insulin resistance in adipose tissue

doi: 10.1111/jdi.13412

Figure Lengend Snippet: Primer used for quantitative polymerase chain reaction analysis

Article Snippet: Thereafter, the tissue slides were then incubated at 4°C overnight with the following primary antibodies: Gas6 (G307; Bioworld) and adiponectin (AHP1005; AbD Serotec, Hercules, CA, USA).

Techniques: Real-time Polymerase Chain Reaction, Sequencing

Journal: Journal of Diabetes Investigation

Article Title: Growth arrest‐specific 6 modulates adiponectin expression and insulin resistance in adipose tissue

doi: 10.1111/jdi.13412

Figure Lengend Snippet: Growth arrest‐specific 6 (Gas6) modulated the expression of adiponectin in adipocytes. (a) The efficiency of Gas6 knockdown in day‐7 (D7) adipocytes. (b) The adipocytokines gene expression (leptin, adiponectin, resistin and retinol‐binding protein 4 [RBP4]) after knockdown Gas6 in D7 adipocytes by real‐time polymerase chain reaction. (c) The adiponectin expression after knockdown Gas6 in D7 adipocytes by enzyme‐linked immunosorbent assay. Data are presented as the mean ± standard deviation. * P < 0.05. GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; mRNA, messenger ribonucleic acid.

Article Snippet: Thereafter, the tissue slides were then incubated at 4°C overnight with the following primary antibodies: Gas6 (G307; Bioworld) and adiponectin (AHP1005; AbD Serotec, Hercules, CA, USA).

Techniques: Expressing, Knockdown, Gene Expression, Binding Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Journal of Diabetes Investigation

Article Title: Growth arrest‐specific 6 modulates adiponectin expression and insulin resistance in adipose tissue

doi: 10.1111/jdi.13412

Figure Lengend Snippet: The expression of growth arrest‐specific 6 (Gas6) and adiponectin in insulin‐resistant obese mice. Mice fed a normal diet (ND; 10% of calories from fat) or high‐fat diet (HF; 60% of calories from fat) for 16 weeks, and blood samples were collected for measurement of blood glucose and insulin. (a) Bodyweight, fasting blood glucose, homeostasis model of assessment of insulin resistance and triglycerides levels are shown. (b) Immunohistochemistry images of Gas6 and adiponectin in adipocyte tissue (magnification: ×40). Data are presented as the mean ± standard deviation. * P < 0.05; n = 10 animals per group.

Article Snippet: Thereafter, the tissue slides were then incubated at 4°C overnight with the following primary antibodies: Gas6 (G307; Bioworld) and adiponectin (AHP1005; AbD Serotec, Hercules, CA, USA).

Techniques: Expressing, Immunohistochemistry, Standard Deviation

Journal: Journal of Diabetes Investigation

Article Title: Growth arrest‐specific 6 modulates adiponectin expression and insulin resistance in adipose tissue

doi: 10.1111/jdi.13412

Figure Lengend Snippet: Metformin ameliorated insulin resistance by enhancing the expression of growth arrest‐specific 6 (Gas6). Using thiazolidinedione (TZD) treatment day‐16 (D16) adipocytes treated with metformin (2 mmol/L) for 6 h. (a) The protein of Gas6, phospho‐adenosine monophosphate‐activated protein kinase (p‐AMPK), adenosine monophosphate‐activated protein kinase (AMPK), phospho‐insulin receptor substrate‐1 (p‐IRS1[307]), insulin receptor substrate‐1(IRS1), phospho‐protein kinase B (p‐Akt) and protein kinase B (Akt) expression by western blot. (b) The adipocytokine gene expressions (adiponectin, leptin, resistin and retinol‐binding protein 4 [RBP4]) by real‐time polymerase chain reaction. (c) The adiponectin expression after metformin treatment in day‐16 (D16) adipocyte by enzyme‐linked immunosorbent assay. Data were presented as the mean ± standard deviation. * P < 0.05. GAPDH, glyceraldehyde 3‐phosphate dehydrogenase.

Article Snippet: Thereafter, the tissue slides were then incubated at 4°C overnight with the following primary antibodies: Gas6 (G307; Bioworld) and adiponectin (AHP1005; AbD Serotec, Hercules, CA, USA).

Techniques: Expressing, Western Blot, Binding Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Drug Design, Development and Therapy

Article Title: 5-( Bis (3-(2-hydroxyethyl)-1H-indol-2-yl)methyl)-2-hydroxybenzoic acid (BHIMHA): showing a strategy of designing drug to block lung metastasis of tumors

doi: 10.2147/DDDT.S93570

Figure Lengend Snippet: Docking investigation of BHIMHA and staurosporine toward PKCα (PDB: 3IW4). Notes: ( A ) The feature of BHIMHA in the active site of PKCα, and the amplified interactions of energy minimized BHIMHA with the amino acid residues. ( B ) The feature of staurosporine in the active site of PKCα, and the amplified interactions of energy minimized staurosporine with the amino acid residues. Abbreviations: PKC, protein kinase C; BHIMHA, 5-( bis (3-(2-hydroxyethyl)-1H-indol-2-yl)-methyl)-2-hydroxybenzoic acid; staurosporine, (5S,6R,7R,9R)-6-methoxy-5-methyl-7-methylamino-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5,9-methanodibenzo[ b, h ]cyclone-na[ jkl ]cyclopenta[ e ]asindacen-14-one; PDB, Protein Data Bank.

Article Snippet: PKCα, PKCβII, PKCγ, PKCη, and PKCδ antibodies were purchased from Boster Biological Engineering Co., Ltd. (Wuhan, People’s Republic of China); PKCθ antibody was purchased from CST (Shanghai) Biological Reagents Company Limited, Shanghai, People’s Republic of China.

Techniques: Amplification

Journal: International Journal of Molecular Sciences

Article Title: Prolonged Chronic Consumption of a High Fat with Sucrose Diet Alters the Morphology of the Small Intestine

doi: 10.3390/ijms22147280

Figure Lengend Snippet: ( A , C ) Immunohistochemistry of adiponectin and adiponectin receptor of proximal tract of the small intestine. (Original magnification: 10×; scale bar: 100 μm). ( B , D ) The expression of adiponectin and its receptor was significantly reduced in HFD w/Suc mice compared to the SD group. * p < 0.05. These images are representative of n = 5 SD, n = 10 HFD w/Suc mice.

Article Snippet: Adiponectin , Boster Biological Technology, Pleasanton, CA, USA; code PA2014-1 , 1:50.

Techniques: Immunohistochemistry, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Prolonged Chronic Consumption of a High Fat with Sucrose Diet Alters the Morphology of the Small Intestine

doi: 10.3390/ijms22147280

Figure Lengend Snippet: Antibodies used with their sources and dilutions.

Article Snippet: Adiponectin , Boster Biological Technology, Pleasanton, CA, USA; code PA2014-1 , 1:50.

Techniques:

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Detection of serum adiponectin isoforms with monoclonal antibodies (mAbs) against recombinant human adiponectin. a Western blot to screen mAbs detecting different adiponectin isoforms. Human serum was loaded into each lane of the gel and resolved by polyacrylamide gel electrophoresis (PAGE) and then transferred to a membrane. Each lane of the membrane was then separately cut and incubated with each mAb from hybridoma culture supernatant. After the secondary antibody was probed, the cut membrane was combined to form one sheet and developed with ECL solution. Eleven different mAbs from culture supernatant were tested to investigate their recognition patterns of adiponectin isoforms in serum. The mAb KH7–33, in lane 3, was selected as a representative recognizing only the middle molecular weight (MMW) isoform of adiponectin. The mAb KH7–41, in lane 4, was selected as a representative recognizing both the MMW and low molecular weight (LMW) isoforms of adiponectin. The mAb KH4–8, in lane 8, was selected as a representative mAb recognizing both the MMW and HMW isoforms of adiponectin. b Comparison of the mAb recognition patterns of adiponectin isoforms in human, mouse, and rat sera by Western blot. Human, mouse, and rat sera were separated by sodium dodecyl sulfate–PAGE. All three mAbs (KH7–33, KH7–41, and KH4–8) recognized the rat and mouse adiponectin MMW isoform. Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011). Mouse and rat serum were obtained via heart puncture of male BALB/c mice and Sprague Dawley rats (8 weeks old), respectively. Human serum was obtained from a male volunteer (55 years old)

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Bioprocessing, Recombinant, Western Blot, Polyacrylamide Gel Electrophoresis, Membrane, Incubation, Molecular Weight, Comparison

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Immunohistological assays in human tissues with monoclonal antibodies (mAbs). To determine the pattern of recognition of adiponectin isoforms in human tissues by mAbs, a normal human adipose tissue was immunostained with mAbs (KH7–41, KH7–33, and KH4–8) (200×, scale bar = 25 μm). mAbs recognized adiponectin in the nucleus of adipocytes (blue arrow) and in vessels and endothelial cells (red arrow). b Human lung, kidney, and pancreas were stained with the KH7–41, KH7–33, and KH4–8 mAbs (100×). Abbreviations: HMW high molecular weight, LMW low molecular weight, MMW middle molecular weight

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Bioprocessing, Staining, High Molecular Weight, Molecular Weight

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Inhibition of adiponectin-mediated gene expression in vitro by monoclonal antibodies (mAbs). To test the ability of the mAb KH4–8 to block adiponectin function, ( a ) human osteoblasts and ( b ) human umbilical vein endothelial cells (HUVECs) were treated with adiponectin (ADIPO) or KH4–8 mAb (mAb) or both. The mAb (~120 μg/mL) and recombinant adiponectin (2.5 μg/mL) were mixed and incubated for 1 h before being used to treat cells. After 24-h treatment, the culture supernatants were collected and frozen, and interleukin-6 (IL-6) and IL-8 were measured by using enzyme-linked immunosorbent assay (ELISA) (R&D Systems, Minneapolis, MN, USA). The experiments were performed in quadruplicate. The data shown are representative of three independent experiments, and similar results were obtained with all three mAbs. Values are expressed as mean ± standard error of the mean. The expression levels of the factors were compared between groups by using the Mann–Whitney test. * P <0.05, ** P <0.01 versus the untreated group, # P <0.05, ## P <0.01 versus the group treated with adiponectin and mAb

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Inhibition, Gene Expression, In Vitro, Bioprocessing, Blocking Assay, Recombinant, Incubation, Enzyme-linked Immunosorbent Assay, Expressing, MANN-WHITNEY

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Epitope mapping of monoclonal antibody (mAb) KH4–8 against adiponectin. To identify the epitope-recognizing site of the KH4–8 mAb, PEPperMAP ® technology was performed as described in the Methods. Human adiponectin was translated into linear 15–amino acid peptides with a peptide-peptide overlap of 14 amino acids. Human adiponectin peptide microarrays were incubated with mouse mAb KH4–8 at different concentrations followed by staining with secondary goat anti-mouse IgG (H + L) DyLight680 antibody. The light intensity was read by a reader. The amino acid sequence QQNHYD (139–144) was confirmed from among the full 244–amino acid sequence to be the epitope of adiponectin

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Incubation, Staining, Sequencing

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Anti-inflammatory effect of monoclonal antibodies (mAbs) KH4–8 and KH7–33 on the expression of serum pro-inflammatory cytokines of the collagen-induced arthritis mouse model. The serum levels of adiponectin, interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and receptor activator of nuclear factor-kappa Β ligand (RANKL) were analyzed by using the Luminex system. Values are expressed as mean ± standard error of the mean. The expression levels of the factors were compared between groups ( n = 8) by using the Mann–Whitney test. ** P <0.01, * P <0.05 versus normal (NOR) group, and ## P <0.01, ## P <0.05 versus the control (CON) group. Abbreviations: ns not significant, pre prednisolone

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Bioprocessing, Expressing, Luminex, MANN-WHITNEY, Control

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Anti-adiponectin antibodies reduce the histological signs of inflammation. The upper and lower panels present hematoxylin and eosin (H&E) staining and immunostaining against mouse adiponectin of mouse knee joints (n = 8), respectively. a Normal, b control, saline-treated arthritic, c KH7–33-treated arthritic, d KH4–8-treated arthritic, and ( e ) prednisolone-treated arthritic mice. Tissue structure was visualized by using H&E staining (original magnification, 40×). Scale bar = 2 mm. f Arthritic symptoms were evaluated by scoring the degree of inflammation on H&E histological sections of knee joints as described in the Methods. Small blue squares on H&E staining are magnified in the upper right corner (400×). Abbreviations: C cartilage, F femur, M meniscus, S subchondral bone, T tibia. In the lower panel, immunohistochemistry (IHC) reveals adiponectin expression in collagen-induced arthritis mouse joints (200×). The increased adiponectin expression level observed on IHC was not decreased by monoclonal antibody treatment. Adiponectin immunostaining score level was evaluated as described in the Methods. Results are presented as the mean of experiments (± standard error of the mean indicated by error bar) (one-way analysis of variance followed by Dunn’s multiple comparison test). *** P <0.001 versus the normal (NOR) group and # P <0.05, ## P <0.01 versus the control (CON) group. Abbreviation: pre prednisolone

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Staining, Immunostaining, Control, Saline, Immunohistochemistry, Expressing, Comparison

Journal: Frontiers in Cell and Developmental Biology

Article Title: Paracrine Effects of Recombinant Human Adiponectin Promote Bone Regeneration

doi: 10.3389/fcell.2021.762335

Figure Lengend Snippet: Mechanisms for local medulla injection of ADPN promoting bone healing. (A) Serum Osteoprotegerin (OPG) levels at weeks 2, 4, and 6. (B) WB analysis of alkaline phosphatase (ALP), bone morphogenic protein 2 (BMP-2), osteocalcin (OCN), and adiponectin receptor 1 (AdipoR1) expressions with corresponding quantification, (C) Immunofluorescent staining of ALP, OCN, BMP-2, and AdipoR1 in G2 (ADPN 1 mg/kg) and G3 (ADPN 2 mg/kg), and the corresponding quantification, n = 5. The white arrow indicates the periosteum, and the blue arrow indicates the lacuna, scale bar = 100 μm.

Article Snippet: Antibodies used for immunofluorescent staining were BMP-2 ab6285 (Abcam, Cambridge, United Kingdom); ALP sc-271431, OCN sc-390877 (Santa Cruz, CA, United States); and AdipoR1 BM4566 (Boster Bio, CA, United States).

Techniques: Injection, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Paracrine Effects of Recombinant Human Adiponectin Promote Bone Regeneration

doi: 10.3389/fcell.2021.762335

Figure Lengend Snippet: The immunofluorescent staining of ALP, OCN, BMP-2, and AdipoR1 on BMSCs with ADPN 10 μg/ml and AdipoR1 siRNA + ADPN 10 μg/ml, and the corresponding quantification, scale bar = 100 μm.

Article Snippet: Antibodies used for immunofluorescent staining were BMP-2 ab6285 (Abcam, Cambridge, United Kingdom); ALP sc-271431, OCN sc-390877 (Santa Cruz, CA, United States); and AdipoR1 BM4566 (Boster Bio, CA, United States).

Techniques: Staining

Journal: Journal of Endocrinology

Article Title: Local adiponectin treatment reduces atherosclerotic plaque size in rabbits

doi: 10.1677/joe-06-0173

Figure Lengend Snippet: Figure 1 Serum adiponectin concentrations in rabbits. 1, Aorta non-injured rabbits on normal cholesterol diet; 2, aorta injured rabbits on high-cholesterol diet before adenovirus transfer; 3, after transfer Ad-bgal through intima; 4, after transfer Ad-APN through intima; 5, after transfer Ad-bgal through adventitia; 6, After transfer Ad-APN through adventitia. The symbol * indicates significant difference (P!0.01) vs serum adiponectin concentrations in aorta non-injured rabbits on normal cholesterol diet; † compared with rabbits before adenovirus transfer; and ‡ compared with rabbits after Ad-bgal transfer. The values are meansGS.E.M.

Article Snippet: For immunohistochemical analyses, paraffin-imbedded cross-sections (5 mm thick) were incubated with either rabbit adiponectin polyclonal antibody (Boster), mouse vascular adhesion molecule-1 (VCAM-1) monoclonal antibody (Santa Cruz), mouse intercellular adhesion molecule-1 (ICAM-1) monoclonal antibody (Santa Cruz), mouse collagen I (Abcam, Cambridge, MA, USA), or mouse collagen III (Calbiochem, Darmstadt, Germany) monoclonal antibody.

Techniques:

Journal: Journal of Endocrinology

Article Title: Local adiponectin treatment reduces atherosclerotic plaque size in rabbits

doi: 10.1677/joe-06-0173

Figure Lengend Snippet: Figure 2 SDS-PAGE (A) and western blot analysis of the supernatant of cultured Pichia pastoris (B, using anti-His tag monoclonal antibody; C, using adiponectin polyclonal antibody). Line 1 is a blank pGAPZa Pichia vector (not containing adiponectin gene) sample control and lines 2–4 are samples from the supernatant of cultured Pichia pastoris containing the recombinant rabbit adiponectin gene. Line 5 contained the molecular weight markers (not visible in image of blot). Both anti-His antibody and adiponectin polyclonal antibody detected a single predominant protein at z42 kDa (containing 9.3 kDa a-factor signal peptide and 2.5 kDa C-terminal His-tag).

Article Snippet: For immunohistochemical analyses, paraffin-imbedded cross-sections (5 mm thick) were incubated with either rabbit adiponectin polyclonal antibody (Boster), mouse vascular adhesion molecule-1 (VCAM-1) monoclonal antibody (Santa Cruz), mouse intercellular adhesion molecule-1 (ICAM-1) monoclonal antibody (Santa Cruz), mouse collagen I (Abcam, Cambridge, MA, USA), or mouse collagen III (Calbiochem, Darmstadt, Germany) monoclonal antibody.

Techniques: SDS Page, Western Blot, Cell Culture, Plasmid Preparation, Control, Recombinant, Molecular Weight

Journal: Journal of Endocrinology

Article Title: Local adiponectin treatment reduces atherosclerotic plaque size in rabbits

doi: 10.1677/joe-06-0173

Figure Lengend Snippet: Figure 5 Effect of Ad-APN treatment on adhesion molecules in atherosclerotic rabbits 14 days after recombinant adenovirus transfer. A, Immunohistochemical detection of adiponectin, VCAM-1 and ICAM-1 in atherosclerotic lesions. Representative images are shown. 1, Ad-bgal transfer through intima. 2, Ad-APN transfer through intima. 3, Ad-bgal transfer through adventitia. 4, Ad-APN transfer through adventitia. The magnification is 400. B, mRNA expression of VCAM-1 and ICAM-1 in abdominal aortic tissue in Ad-bgal or Ad-APN-treated rabbits. The values are meansGS.E.M.

Article Snippet: For immunohistochemical analyses, paraffin-imbedded cross-sections (5 mm thick) were incubated with either rabbit adiponectin polyclonal antibody (Boster), mouse vascular adhesion molecule-1 (VCAM-1) monoclonal antibody (Santa Cruz), mouse intercellular adhesion molecule-1 (ICAM-1) monoclonal antibody (Santa Cruz), mouse collagen I (Abcam, Cambridge, MA, USA), or mouse collagen III (Calbiochem, Darmstadt, Germany) monoclonal antibody.

Techniques: Recombinant, Immunohistochemical staining, Expressing

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Detection of serum adiponectin isoforms with monoclonal antibodies (mAbs) against recombinant human adiponectin. a Western blot to screen mAbs detecting different adiponectin isoforms. Human serum was loaded into each lane of the gel and resolved by polyacrylamide gel electrophoresis (PAGE) and then transferred to a membrane. Each lane of the membrane was then separately cut and incubated with each mAb from hybridoma culture supernatant. After the secondary antibody was probed, the cut membrane was combined to form one sheet and developed with ECL solution. Eleven different mAbs from culture supernatant were tested to investigate their recognition patterns of adiponectin isoforms in serum. The mAb KH7–33, in lane 3, was selected as a representative recognizing only the middle molecular weight (MMW) isoform of adiponectin. The mAb KH7–41, in lane 4, was selected as a representative recognizing both the MMW and low molecular weight (LMW) isoforms of adiponectin. The mAb KH4–8, in lane 8, was selected as a representative mAb recognizing both the MMW and HMW isoforms of adiponectin. b Comparison of the mAb recognition patterns of adiponectin isoforms in human, mouse, and rat sera by Western blot. Human, mouse, and rat sera were separated by sodium dodecyl sulfate–PAGE. All three mAbs (KH7–33, KH7–41, and KH4–8) recognized the rat and mouse adiponectin MMW isoform. Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011). Mouse and rat serum were obtained via heart puncture of male BALB/c mice and Sprague Dawley rats (8 weeks old), respectively. Human serum was obtained from a male volunteer (55 years old)

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Bioprocessing, Recombinant, Western Blot, Polyacrylamide Gel Electrophoresis, Membrane, Incubation, Molecular Weight, Comparison

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Immunohistological assays in human tissues with monoclonal antibodies (mAbs). To determine the pattern of recognition of adiponectin isoforms in human tissues by mAbs, a normal human adipose tissue was immunostained with mAbs (KH7–41, KH7–33, and KH4–8) (200×, scale bar = 25 μm). mAbs recognized adiponectin in the nucleus of adipocytes (blue arrow) and in vessels and endothelial cells (red arrow). b Human lung, kidney, and pancreas were stained with the KH7–41, KH7–33, and KH4–8 mAbs (100×). Abbreviations: HMW high molecular weight, LMW low molecular weight, MMW middle molecular weight

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Bioprocessing, Staining, High Molecular Weight, Molecular Weight

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Inhibition of adiponectin-mediated gene expression in vitro by monoclonal antibodies (mAbs). To test the ability of the mAb KH4–8 to block adiponectin function, ( a ) human osteoblasts and ( b ) human umbilical vein endothelial cells (HUVECs) were treated with adiponectin (ADIPO) or KH4–8 mAb (mAb) or both. The mAb (~120 μg/mL) and recombinant adiponectin (2.5 μg/mL) were mixed and incubated for 1 h before being used to treat cells. After 24-h treatment, the culture supernatants were collected and frozen, and interleukin-6 (IL-6) and IL-8 were measured by using enzyme-linked immunosorbent assay (ELISA) (R&D Systems, Minneapolis, MN, USA). The experiments were performed in quadruplicate. The data shown are representative of three independent experiments, and similar results were obtained with all three mAbs. Values are expressed as mean ± standard error of the mean. The expression levels of the factors were compared between groups by using the Mann–Whitney test. * P <0.05, ** P <0.01 versus the untreated group, # P <0.05, ## P <0.01 versus the group treated with adiponectin and mAb

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Inhibition, Gene Expression, In Vitro, Bioprocessing, Blocking Assay, Recombinant, Incubation, Enzyme-linked Immunosorbent Assay, Expressing, MANN-WHITNEY

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Epitope mapping of monoclonal antibody (mAb) KH4–8 against adiponectin. To identify the epitope-recognizing site of the KH4–8 mAb, PEPperMAP ® technology was performed as described in the Methods. Human adiponectin was translated into linear 15–amino acid peptides with a peptide-peptide overlap of 14 amino acids. Human adiponectin peptide microarrays were incubated with mouse mAb KH4–8 at different concentrations followed by staining with secondary goat anti-mouse IgG (H + L) DyLight680 antibody. The light intensity was read by a reader. The amino acid sequence QQNHYD (139–144) was confirmed from among the full 244–amino acid sequence to be the epitope of adiponectin

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Incubation, Staining, Sequencing

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Anti-inflammatory effect of monoclonal antibodies (mAbs) KH4–8 and KH7–33 on the expression of serum pro-inflammatory cytokines of the collagen-induced arthritis mouse model. The serum levels of adiponectin, interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and receptor activator of nuclear factor-kappa Β ligand (RANKL) were analyzed by using the Luminex system. Values are expressed as mean ± standard error of the mean. The expression levels of the factors were compared between groups ( n = 8) by using the Mann–Whitney test. ** P <0.01, * P <0.05 versus normal (NOR) group, and ## P <0.01, ## P <0.05 versus the control (CON) group. Abbreviations: ns not significant, pre prednisolone

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Bioprocessing, Expressing, Luminex, MANN-WHITNEY, Control

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Anti-adiponectin antibodies reduce the histological signs of inflammation. The upper and lower panels present hematoxylin and eosin (H&E) staining and immunostaining against mouse adiponectin of mouse knee joints (n = 8), respectively. a Normal, b control, saline-treated arthritic, c KH7–33-treated arthritic, d KH4–8-treated arthritic, and ( e ) prednisolone-treated arthritic mice. Tissue structure was visualized by using H&E staining (original magnification, 40×). Scale bar = 2 mm. f Arthritic symptoms were evaluated by scoring the degree of inflammation on H&E histological sections of knee joints as described in the Methods. Small blue squares on H&E staining are magnified in the upper right corner (400×). Abbreviations: C cartilage, F femur, M meniscus, S subchondral bone, T tibia. In the lower panel, immunohistochemistry (IHC) reveals adiponectin expression in collagen-induced arthritis mouse joints (200×). The increased adiponectin expression level observed on IHC was not decreased by monoclonal antibody treatment. Adiponectin immunostaining score level was evaluated as described in the Methods. Results are presented as the mean of experiments (± standard error of the mean indicated by error bar) (one-way analysis of variance followed by Dunn’s multiple comparison test). *** P <0.001 versus the normal (NOR) group and # P <0.05, ## P <0.01 versus the control (CON) group. Abbreviation: pre prednisolone

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Staining, Immunostaining, Control, Saline, Immunohistochemistry, Expressing, Comparison

Journal: PLoS ONE

Article Title: Mechanism of Butyrate Stimulation of Triglyceride Storage and Adipokine Expression during Adipogenic Differentiation of Porcine Stromovascular Cells

doi: 10.1371/journal.pone.0145940

Figure Lengend Snippet: (A) Representative immunoblotting and quantification analysis of adiponectin abundance in culture media. (B) Representative immunoblotting analysis of intracellular adiponectin level in the cell lysate. Preadipocytes were treated with butyrate at 1500 μM concentration throughout the differentiation period (day 0 to 9). Both cells and culture media were harvested on day 9 for immunoblotting. Data were analyzed by t test, and P values were shown on the bars (Control vs. Butyrate). Data are means ± SE (n = 4).

Article Snippet: The following primary antibodies were used: anti-PPARγ, anti-phospho Akt, anti-Akt, anti-phospho AMPKα, anti-AMPKα, anti-phospho ACC, anti-ACC, anti-β-actin (Cell Signaling Technology, Danvers, MA, USA), anti-DGAT1 (Neobiolab, Cambridge MA, USA), anti-DGAT2 (Abcam, Cambridge MA, USA), anti-adiponectin (Xeno Diagnostics, LLC, IN, USA) and anti-albumin (BioVision, Inc., Milpitas, CA, USA).

Techniques: Western Blot, Concentration Assay

Journal: PLoS ONE

Article Title: Mechanism of Butyrate Stimulation of Triglyceride Storage and Adipokine Expression during Adipogenic Differentiation of Porcine Stromovascular Cells

doi: 10.1371/journal.pone.0145940

Figure Lengend Snippet: Lines with arrowhead represent upregulation of activity, protein content, and/or expression. Lines without arrowhead denote inhibition. Butyrate supplementation leads to induction of adipogenesis, which results in increased adiponectin secretion. Adiponectin activates the AKT pathway, thereby increasing insulin sensitivity. Adiponectin also leads to activation of AMPK. Butyrate increases intracellular triglyceride accumulation through inhibition of lipolysis and enhancement of triglyceride synthesis. Butyrate also inhibits peroxisomal fatty acid oxidation, leading to accumulation of LCFAs, which may also account for feedback inhibition of de novo fatty acid synthesis through increased ACC phosphorylation.

Article Snippet: The following primary antibodies were used: anti-PPARγ, anti-phospho Akt, anti-Akt, anti-phospho AMPKα, anti-AMPKα, anti-phospho ACC, anti-ACC, anti-β-actin (Cell Signaling Technology, Danvers, MA, USA), anti-DGAT1 (Neobiolab, Cambridge MA, USA), anti-DGAT2 (Abcam, Cambridge MA, USA), anti-adiponectin (Xeno Diagnostics, LLC, IN, USA) and anti-albumin (BioVision, Inc., Milpitas, CA, USA).

Techniques: Activity Assay, Expressing, Inhibition, Activation Assay