Journal: Current obesity reports

Article Title: FSP27 and links to obesity and diabetes mellitus

doi: 10.1007/s13679-019-00343-3

Figure Lengend Snippet: Summary of cellular studies and murine models

Article Snippet: J. Obes Humans: Lean and obese Low FSP27 in obesity vs. lean, low FSP27 in visceral than in subcutaneous fat, negatively correlated with blood parameters Grahn et. al. 2014 J Biol Chem Human adipocytes: Silencing/overexpression of FSP27 Silencing of FSP27 impaired insulin signaling, overexpression protected from FFA induced insulin resistance Sharma et. al. 2018 Bioscientifica Murine adipocyte: overexpression of FSP27 Protects from growth hormone induced insulin resistance Sharma et. al. 2019 Am J Physiol Endocrinol Metab Healthy humans: growth hormones/saline infusion Growth hormone mediated reduction in FSP27, increased lipolysis and FFA, insulin resistance Open in a separate window Summary human studies

Techniques: In Vivo, Knockdown, Control, Over Expression

Journal: Current obesity reports

Article Title: FSP27 and links to obesity and diabetes mellitus

doi: 10.1007/s13679-019-00343-3

Figure Lengend Snippet: Summary human studies

Article Snippet: J. Obes Humans: Lean and obese Low FSP27 in obesity vs. lean, low FSP27 in visceral than in subcutaneous fat, negatively correlated with blood parameters Grahn et. al. 2014 J Biol Chem Human adipocytes: Silencing/overexpression of FSP27 Silencing of FSP27 impaired insulin signaling, overexpression protected from FFA induced insulin resistance Sharma et. al. 2018 Bioscientifica Murine adipocyte: overexpression of FSP27 Protects from growth hormone induced insulin resistance Sharma et. al. 2019 Am J Physiol Endocrinol Metab Healthy humans: growth hormones/saline infusion Growth hormone mediated reduction in FSP27, increased lipolysis and FFA, insulin resistance Open in a separate window Summary human studies

Techniques: Mutagenesis, Over Expression

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Fat‐Specific Protein 27 Regulation of Vascular Function in Human Obesity

doi: 10.1161/JAHA.118.011431

Figure Lengend Snippet: Comparison of FSP 27 (fat‐specific protein 27) protein in primary endothelial cells from subcutaneous vs visceral fat of obese subjects. A , Representative immunofluorescence image of isolated endothelial cells demonstrating significantly reduced expression of FSP 27 protein (red= FSP 27, green= CD 31, endothelial cell marker, blue=DAPI, a nuclear stain) in endothelial cells isolated from visceral compared with subcutaneous depots from obese subjects. B , Quantification of FSP 27 protein expression in endothelial cells isolated from subcutaneous and visceral fat (n=14, P <0.01). Data are presented as arbitrary units and as mean± SEM , indexed to 1 for the subcutaneous depot. *indicates statistical significance, such as P < 0.05.

Article Snippet: Proteins of interests were FSP27 (Catalogue #PA5‐30793, Thermofisher); p‐eNOS at serine 1177 (Catalogue #ab184154, Abcam); pAKT (protein kinase B) at serine 473 (Catalogue #44‐609G, Thermofisher); VEGF‐A (Catalogue #PA5‐16754, Thermofisher), and GAPDH as a loading control.

Techniques: Comparison, Immunofluorescence, Isolation, Expressing, Marker, Staining

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Fat‐Specific Protein 27 Regulation of Vascular Function in Human Obesity

doi: 10.1161/JAHA.118.011431

Figure Lengend Snippet: Insulin‐mediated activation of eNOS (endothelial NO synthase) and AKT (protein Kinase B) in response to recombinant FSP 27 in visceral depot. A , Representative visceral adipose tissue immunoblot demonstrating severe impairment in insulin‐mediated activation of eNOS and AKT in visceral fat. After 24 hours of rFSP 27 (recombinant FSP 27) exposure, insulin‐mediated activation is restored. B , Quantification of percent change in insulin‐mediated activation of eNOS at baseline and after 24 hours of treatment with rFSP 27 in the visceral depot. C , Quantification of percent change in insulin‐mediated activation of AKT at baseline and after 24 hours of treatment with rFSP 27 in the visceral depot (n=10, P <0.05). Data are presented as arbitrary units (au) and as mean±SEM. rFSP27 indicates recombinant fat‐specific protein 27. *indicates statistical significance, such as P < 0.05.

Article Snippet: Proteins of interests were FSP27 (Catalogue #PA5‐30793, Thermofisher); p‐eNOS at serine 1177 (Catalogue #ab184154, Abcam); pAKT (protein kinase B) at serine 473 (Catalogue #44‐609G, Thermofisher); VEGF‐A (Catalogue #PA5‐16754, Thermofisher), and GAPDH as a loading control.

Techniques: Activation Assay, Recombinant, Western Blot

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Fat‐Specific Protein 27 Regulation of Vascular Function in Human Obesity

doi: 10.1161/JAHA.118.011431

Figure Lengend Snippet: Effect of FSP 27 overexpression of FSP 27 and insulin on nitric oxide ( NO ) production in primary endothelial cells isolated from visceral fat. A , Human adenovirus construct of FSP 27 and control adenovirus was transfected into primary endothelial cells isolated from visceral fat (n=5, P <0.01). B , Quantification of insulin‐mediated production of NO in control virus vs FSP 27 over‐expressed primary endothelial cells isolated from visceral fat (n=5, P <0.05). Data are presented as arbitrary units (au) and as mean± SEM . FSP27 indicates fat‐specific protein 27. *indicates statistical significance, such as P < 0.05.

Article Snippet: Proteins of interests were FSP27 (Catalogue #PA5‐30793, Thermofisher); p‐eNOS at serine 1177 (Catalogue #ab184154, Abcam); pAKT (protein kinase B) at serine 473 (Catalogue #44‐609G, Thermofisher); VEGF‐A (Catalogue #PA5‐16754, Thermofisher), and GAPDH as a loading control.

Techniques: Over Expression, Isolation, Construct, Control, Transfection, Virus

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Fat‐Specific Protein 27 Regulation of Vascular Function in Human Obesity

doi: 10.1161/JAHA.118.011431

Figure Lengend Snippet: Insulin‐mediated activation of eNOS (endothelial NO synthase) and AKT (protein Kinase B) in response to si RNA ‐mediated knockdown of FSP 27 in the subcutaneous depot. A , Representative immunoblot demonstrating insulin‐mediated activation of eNOS and AKT in subcutaneous fat under scrambled si RNA (small interfering RNA ) conditions and after knockdown of FSP 27 by si RNA . B , Quantification of percent change in insulin‐mediated activation of eNOS at baseline and after si RNA ‐mediated knockdown of FSP 27 in the subcutaneous fat depot. C , Quantification of percent change in insulin‐mediated activation of AKT at baseline and after si RNA ‐mediated knockdown of FSP 27 in subcutaneous fat depot (n=10, P <0.05). Data are presented as arbitrary units and as mean±SEM. FSP27 indicates fat‐specific protein 27. *indicates statistical significance, such as P < 0.05.

Article Snippet: Proteins of interests were FSP27 (Catalogue #PA5‐30793, Thermofisher); p‐eNOS at serine 1177 (Catalogue #ab184154, Abcam); pAKT (protein kinase B) at serine 473 (Catalogue #44‐609G, Thermofisher); VEGF‐A (Catalogue #PA5‐16754, Thermofisher), and GAPDH as a loading control.

Techniques: Activation Assay, Knockdown, Western Blot, Small Interfering RNA

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Fat‐Specific Protein 27 Regulation of Vascular Function in Human Obesity

doi: 10.1161/JAHA.118.011431

Figure Lengend Snippet: In‐vitro angiogenic capacity of primary endothelial cells from visceral fat in response to recombinant FSP 27 exposure. A , Representative images of primary endothelial cells isolated from visceral fat that exhibited increased angiogenic branch formation when exposed to rFSP 27 (recombinant FSP 27) compared with control. B , Quantification of in‐vitro angiogenic capacity of primary endothelial, cells with and without rFSP 27 treatment for up to 6 hours (n=7, P <0.05). rFSP27 indicates recombinant fat‐specific protein 27. *indicates statistical significance, such as P < 0.05.

Article Snippet: Proteins of interests were FSP27 (Catalogue #PA5‐30793, Thermofisher); p‐eNOS at serine 1177 (Catalogue #ab184154, Abcam); pAKT (protein kinase B) at serine 473 (Catalogue #44‐609G, Thermofisher); VEGF‐A (Catalogue #PA5‐16754, Thermofisher), and GAPDH as a loading control.

Techniques: In Vitro, Recombinant, Isolation, Control

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Fat‐Specific Protein 27 Regulation of Vascular Function in Human Obesity

doi: 10.1161/JAHA.118.011431

Figure Lengend Snippet: Interaction of FSP 27 and VEGF ‐A. A , Co‐immunoprecipitation of FSP 27 followed by immunoblot of VEGF ‐A in cultured human aortic endothelial cells demonstrating an interaction between FSP 27 and VEGF ‐A. B , In‐vitro transcription translation of FSP 27 and VEGF ‐A followed by immunoblot of FSP 27 depicting an interaction between FSP 27 and VEGF ‐A. C , Western blot showing that si RNA (small interfering RNA )‐mediated inhibition of FSP 27 nearly fully abolished VEGF ‐A in human aortic endothelial cells. Experiments were repeated 3 times. FSP27 indicates fat‐specific protein 27; IP, immunoprecipitation; IB, immunoblot; VEFG‐A, vascular endothelial growth factor‐A.

Article Snippet: Proteins of interests were FSP27 (Catalogue #PA5‐30793, Thermofisher); p‐eNOS at serine 1177 (Catalogue #ab184154, Abcam); pAKT (protein kinase B) at serine 473 (Catalogue #44‐609G, Thermofisher); VEGF‐A (Catalogue #PA5‐16754, Thermofisher), and GAPDH as a loading control.

Techniques: Immunoprecipitation, Western Blot, Cell Culture, In Vitro, Small Interfering RNA, Inhibition

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Fat‐Specific Protein 27 Regulation of Vascular Function in Human Obesity

doi: 10.1161/JAHA.118.011431

Figure Lengend Snippet: Angiogenic capacity and FSP 27 mRNA expression of subcutaneous fat after weight‐loss. A , Longitudinally, paired fat tissue explants demonstrated increased angiogenic growth in subcutaneous depots after weight‐loss ( B ) mRNA expression of FSP 27 increased following weight loss and ( C ) positively correlated with angiogenic sprouting (n=7, P <0.05). au indicates arbitrary units; FSP27, fat‐specific protein 27. *indicates statistical significance, such as P < 0.05.

Article Snippet: Proteins of interests were FSP27 (Catalogue #PA5‐30793, Thermofisher); p‐eNOS at serine 1177 (Catalogue #ab184154, Abcam); pAKT (protein kinase B) at serine 473 (Catalogue #44‐609G, Thermofisher); VEGF‐A (Catalogue #PA5‐16754, Thermofisher), and GAPDH as a loading control.

Techniques: Expressing