Journal: Cell Transplantation

Article Title: Comparing Scaffold-Free and Fibrin-Based Adipose-Derived Stromal Cell Constructs for Adipose Tissue Engineering: An In Vitro and in Vivo Study

doi: 10.3727/096368912x653129

Figure Lengend Snippet: Figure 6. ASCs in fibrin-based constructs display increased adipogenic differentiation when compared to ASCs in spheroids. (A) ASCs in spheroids and fibrin-based constructs were cultured for 7 days in vitro in adipogenic medium. Cross-sections were stained with Oil Red O (showing intracellular lipid) and counterstained with haematoxylin. (A.1) Spheroid. (A.2) Fibrin-based construct. Scale bars: 100 µm. (B) Q-PCR was used to measure the expression levels of the adipocyte-specific markers peroxisome proliferator-activated receptor g (PPARG), perilipin 1 (PLIN 1), fatty acid binding protein 4 (FABP4), and leptin (LEP). Expression levels are relative to b-2- microglobulin-positive control housekeeping gene (dCt). Values are presented as medians (interquartile range). For each of the three ASC donors, nine spheroids, nine fibrin-based constructs, and 0.5 ´ 106 undifferentiated ASCs (=control) were prepared and assayed. *p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.001 (relative expression in fibrin-based constructs compared to spheroids and control). (C) The secre- tion of leptin by ASCs in conditioned medium of spheroids and fibrin-based constructs over 48 h was measured by sandwich ELISA at day 7. Unconditioned adipogenic medium was used as control condition. Values are presented as median (interquartile range); n = 9, 3 ASC donors. **p ≤ 0.01, ***p ≤ 0.001 (fibrin-based construct conditioned medium vs. spheroid conditioned medium and control).

Article Snippet: The mRNA levels of the adipogenic markers peroxisome proliferator-activated receptor g (PPARG), fatty acid binding protein 4 (FABP4), perilipin 1 (PLIN 1), and leptin (LEP), together with b-2-microglobulin (B2M), were analyzed with the ABI PRISM® 7000 Sequence Detection System and 7000 System SDS software (ABI, Foster City, CA, USA, www.appliedbiosystems.com), using the Taqman®Gene Expression Assays for PPARG (Hs01115510_m1), FABP4 (Hs01086177_m1), PLIN1 (Hs01106927_m1), LEP (Hs00174877_m1), and B2M (Hs00984230_m1) (ABI) according to the manufacturer's instructions.

Techniques: Construct, Cell Culture, In Vitro, Staining, Expressing, Binding Assay, Positive Control, Control, Sandwich ELISA

Journal: Nutrients

Article Title: Sex-Dependent Mediation of Leptin in the Association of Perilipin Polymorphisms with BMI and Plasma Lipid Levels in Children

doi: 10.3390/nu14153072

Figure Lengend Snippet: Genotypic and allelic distribution of the SNPs studied in PLIN1 and PLIN2 .

Article Snippet: To determine the polymorphism in the perilipin genes, the following predesigned TaqMan SNP Genotyping Assays from Applied Biosystems (Waltham, MA, USA) were used: C_8722593_10, C_8722587_10, and C_9304320_20 for the SNPs in PLIN1 rs894160, rs1052700, and rs2304795, respectively, and C_25764255_10 for the SNPrs35568725 in PLIN2 .

Techniques:

Journal: Nutrients

Article Title: Sex-Dependent Mediation of Leptin in the Association of Perilipin Polymorphisms with BMI and Plasma Lipid Levels in Children

doi: 10.3390/nu14153072

Figure Lengend Snippet: BMI and lipid profile values (means ± SD) by genotype for PLIN1 and PLIN2 SNPs in boys and girls.

Article Snippet: To determine the polymorphism in the perilipin genes, the following predesigned TaqMan SNP Genotyping Assays from Applied Biosystems (Waltham, MA, USA) were used: C_8722593_10, C_8722587_10, and C_9304320_20 for the SNPs in PLIN1 rs894160, rs1052700, and rs2304795, respectively, and C_25764255_10 for the SNPrs35568725 in PLIN2 .

Techniques:

Journal: Nutrients

Article Title: Sex-Dependent Mediation of Leptin in the Association of Perilipin Polymorphisms with BMI and Plasma Lipid Levels in Children

doi: 10.3390/nu14153072

Figure Lengend Snippet: ( a ) BMI values of PLIN1 rs894160 and PLIN2 rs35568725 genotypes in boys and girls according to levels of leptin; ( b ) NEFA levels of PLIN1 rs2304795 and PLIN2 rs35568725 genotypes in boys and girls by leptin levels; ( c ) Apo A-I levels of PLIN1 rs1052700 genotypes in boys and girls by levels of leptin. ( d ) HDL-cholesterol levels of PLIN1 rs1052700 genotypes in boys and girls by levels of leptin. Values are expressed as median and interquartile range. p -value: Mann–Whitney U test: * p -value < 0.05; ** p -value < 0.01.

Article Snippet: To determine the polymorphism in the perilipin genes, the following predesigned TaqMan SNP Genotyping Assays from Applied Biosystems (Waltham, MA, USA) were used: C_8722593_10, C_8722587_10, and C_9304320_20 for the SNPs in PLIN1 rs894160, rs1052700, and rs2304795, respectively, and C_25764255_10 for the SNPrs35568725 in PLIN2 .

Techniques: MANN-WHITNEY

Journal: American journal of physiology. Heart and circulatory physiology

Article Title: Chemerin is resident to vascular tunicas and contributes to vascular tone

doi: 10.1152/ajpheart.00239.2023

Figure Lengend Snippet: A. Detection of chemerin protein expression in tunicas of the thoracic aorta from Dahl SS male and female rats. P = PVAT, A = adventitia; M = media, E = endothelium, L = lumen. The first row demonstrates colocalization of chemerin signal (green) with smooth muscle cell (SMC) a-actin (red) and second row with the adipocyte marker perilipin (red). DAPI (blue) is shown in the leftmost box, for each image. Images are representative of 4/5 different rats. B. Detection of chemerin protein in the human epigastric artery. Images are representative of 3 different human samples. Negative controls shown in bottom left corner for each image. 20x images, scale bar = 100 μm

Article Snippet: Sections were then species-specific blocked with protein blocker for one hour, followed by an incubation for 24 hours with chemerin (cat. #H-002–52, rabbit TIG-2; 1:200, Phoenix Pharmaceuticals Inc, Belmont, CA, USA), alpha actin (cat. # C6198, alpha actin-Cy3, incubated simultaneously with chemerin,1:1000, Sigma-Aldrich, St. Louis, MO, USA), or perilipin (cat. # AM09128SU-N, incubated sequentially prior to chemerin, ready to use, Origene, Herford, Germany) or no primary antibody at 4 °C in a humidified, closed chamber.

Techniques: Expressing, Marker