No data available that match "3T3-L1 Cells"

*  Antiadipogenic Effect of Dietary Apigenin through Activation of AMPK in 3T3-L1 Cells

Antiadipogenic Effect of Dietary Apigenin through Activation of AMPK in 3T3-L1 Cells ...

*  Unraveling the mode of action of an obesogen : mechanistic analysis of the model obesogen tributyltin in the 3T3-L1 cell line -...

Unraveling the mode of action of an obesogen : mechanistic analysis of the model obesogen tributyltin in the 3T3-L1 cell line ... Furthermore, our results show that combining transcriptomics with 3T3-L1 cells is a promising tool for screening of potential ... This study was designed to unravel the molecular mechanisms of the model obesogen TBT, using microarray analysis in the 3T3-L1 ... combining effects on both cell physiological and gene expression level. ...

*  COL5A3 - Anti-COL5A3 Antibodies, shRNA, siRNA & Gene Information | Sigma-Aldrich

Cell Line: 3T3-L1. TRC Version: 1. Clone ID:NM_016919.1-5890s1c1. ... Validated MISSION® siRNAs have been functionally validated in Hela cells by Sigma scientists to Knockdown gene expression by 75 ...®ion=

*  NOD1 activation induces proinflammatory gene expression and insulin resistance in 3T3-L1 adipocytes | Endocrinology and...

... and adipocyte states have been studied in 3T3-L1 cells and their functionalities demonstrated in 3T3-L1 (25, 31, 32) and human ... NOD1 ligand Tri-DAP suppressed insulin-induced glucose uptake in 3T3-L1 adipocytes. 3T3-L1 adipocytes were starved in Krebs- ... 3T3-L1 adipocytes were stimulated with vehicle or Tri-DAP (10 μg/ml) for indicated periods of time, and whole cell lysates were ... 3T3-L1 cells. obesity is becoming a global epidemic that affects both adults and children. Accumulating evidence indicates that ...

*  JCI - Pyruvate carboxylase is critical for non-small-cell lung cancer proliferation

of biotin-containing proteins in Swiss 3T3-L1 cells. Biochem J. 1988;251(3):749-755.. View this article via: PubMed Google ... PC-knockdown cells had over 70% more basal ROS than did control cells (0 mM H2O2; Supplemental Figure 12D). When cells were ... shRNA knockdown of PC in lung cancer cells. NSCLC cells lines and HEK 293T cells obtained from ATCC were grown in DMEM media ... HCC827 and PC9 cells were more sensitive to PC knockdown than were A549, H2030, or H1299 cells. HCC827 and PC9 cells also ...

*  Active Motif » Cell Extracts, Tissue Extracts, Nuclear, Whole-cell and Cytoplasmic Extracts

... cytoplasmic and whole-cell extracts from a variety of cell types and tissue sources that are ready to use as controls in a ... 3T3-L1 nuclear extract 200 µg 36071 $165 Buy Now A-431 nuclear extract 200 µg 36004 $165 Buy Now ... Hep G2 whole-cell extract 200 µg 36412 $165 Buy Now Hep G2 whole-cell extract (IL-6 stimulated, 10 ng/ml) 200 µg 36413 $165 Buy ... NIH/3T3 whole-cell extract (TPA stimulated) 200 µg 36415 $165 Buy Now ...

*  Alumni | Biochemistry and Molecular Biology | East Carolina University

Dissertation: "Post-Transcriptional Regulation of mRNA Metabolism during Differentiation of 3T3-L1 Cells: Role of HuR" ... Dissertation: "N-3 Polyunsaturated Fatty Acids Differentially Enhance B-cell Mediated Immunity in Lean & Obese Mice" ... Postdoctoral: Christopher Geyer, PhD., Department of Anatomy & Cell Biology, The Brody School of Medicine, East Carolina ... Dissertation: "Fish oil disrupts B cell plasma membrane lateral organization and immunological synapse formation" ...

*  Nutrients | Free Full-Text | Non-Alcoholic Steatohepatitis and Hepatocellular Carcinoma: Implications for Lycopene...

3T3-L1 cells; adipose tissue explants of mice subjected to HFD in ex vivo culture; primo culture of human mature adipocytes in ... Interferons and tumor necrosis factors have similar catabolic effects on 3T3 L1 cells. Proc. Natl. Acad. Sci. USA 1986, 83, ... Of relevance to NAFLD and obesity, it was observed that pretreatment of preadipocytes, differentiated 3T3-L1 adipocytes, and ... TLR4 is expressed in all types of liver cells [130]. KCs are among the first liver cells to be exposed to intestinal-derived ...

*  Evolution of lactation: nutrition v. protection with special reference to five mammalian species | Nutrition Research Reviews |...

2007) Growth hormone stimulates adipogenesis of 3T3-L1 cells through activation of the Stat5A/5B-PPARγ pathway. J Mol ... messenger RNA expression and stimulates adipogenic conversion of NIH-3T3 cells. Mol Endocrinol 14, 307-316. ...

*  stable 3t3l1 transfection:serious help needed!!! - Molecular Biology - BioForum

I m just getting started with stabile transfections of 3t3L1 cells. and i am the only one in my lab to ever do this.i have been ... stable 3t3l1 transfection:serious help needed!!! - posted in Molecular Biology: Hey Guys, ... One thing you should know about 3T3-L1 is that they are pretty, pretty hard to transfect. Even adenovirus, at an MOI of 2000, ... my 3t3L1 cells? i m sure u can probably buy the vector itself with a Antibiotic resistance (thanks for the tip with puromycin ...

No data available that match "3T3-L1 Cells"

(1/1584) Role of cyclooxygenases COX-1 and COX-2 in modulating adipogenesis in 3T3-L1 cells.

Cyclooxygenase (COX) catalyses the rate-limiting step of prostanoid biosynthesis. Two COX isoforms have been identified, COX-1, the constitutive form, and COX-2, the inducible form. While COX-2 has been implicated in body fat regulation, the underlying cellular mechanism remains to be elucidated. The present study was undertaken to examine the potential role of COX in modulating adipogenesis and to dissect the relative contribution of the two isoenzymes in this process. COX-2 was found to be expressed in undifferentiated 3T3-L1 cells and down-regulated during differentiation, whereas the cellular level of COX-1 remained relatively constant. Abrogating the activity of either of these two isoenzymes by selective COX inhibitors accelerated cellular differentiation, suggesting that both COX isoenzymes negatively influenced differentiation. Tumor necrosis factor-alpha (TNFalpha) significantly up-regulated COX-2 expression ( approximately 2-fold) in differentiating 3T3-L1 cells, whereas similar effect was not observed with COX-1 expression. Abrogating the induced COX-2 activity reversed the TNFalpha-induced inhibition of differentiation by approximately 70%, implying a role for COX-2 in mediating TNFalpha signaling. Hence, both COX isoforms were involved in the negative modulation of adipocyte differentiation. COX-2 appeared to be the main isoform mediating at least part of the negative effects of TNFalpha.  (+info)

(2/1584) Isomer-specific regulation of metabolism and PPARgamma signaling by CLA in human preadipocytes.

Trans-10,cis-12 conjugated linoleic acid (CLA) has previously been shown to be the CLA isomer responsible for CLA-induced reductions in body fat in animal models, and we have shown that this isomer, but not the cis-9,trans-11 CLA isomer, specifically decreased triglyceride (TG) accumulation in primary human adipocytes in vitro. Here we investigated the mechanism behind the isomer-specific, CLA-mediated reduction in TG accumulation in differentiating human preadipocytes. Trans-10,cis-12 CLA decreased insulin-stimulated glucose uptake and oxidation, and reduced insulin-dependent glucose transporter 4 gene expression. Furthermore, trans-10,cis-12 CLA reduced oleic acid uptake and oxidation when compared with all other treatments. In parallel to CLA's effects on metabolism, trans-10,cis-12 CLA decreased, whereas cis-9,trans-11 CLA increased, the expression of peroxisome proliferator-activated receptor gamma (PPARgamma) and several of its downstream target genes when compared with vehicle controls. Transient transfections demonstrated that both CLA isomers antagonized ligand-dependent activation of PPARgamma. Collectively, trans-10,cis-12, but not cis-9, trans-11, CLA decreased glucose and lipid uptake and oxidation and preadipocyte differentiation by altering preadipocyte gene transcription in a manner that appeared to be due, in part, to decreased PPARgamma expression.  (+info)

(3/1584) Regulation of ABCA1 expression and cholesterol efflux during adipose differentiation of 3T3-L1 cells.

Adipose cells specialized in energy storage, contain large intracellular triglyceride-rich lipid droplets, are enriched with free cholesterol, and express sterol-regulated transcription factors such as liver X receptor (LXR). The recent identification of the LXR-dependent ATP binding cassette transporter A1 (ABCA1) pathway for cholesterol release from peripheral cells has led us to address the question of the expression and function of ABCA1 in adipocytes. In 3T3-L1 adipose cells, we observed a strong induction of ABCA1 mRNA during adipose differentiation, but only limited variations in ABCA1 protein. Lipid efflux onto apolipoprotein A-I (apoA-I), which depends on ABCA1, was comparable in adipocytes and preadipocytes, demonstrating a differential regulation of ABCA1 mRNA and cholesterol efflux. We also found that total cell cholesterol remained stable during differentiation of 3T3-L1 cells, but membrane cholesterol was lower in adipocytes than in preadipocytes, suggesting redistribution of cholesterol to the lipid droplet. Finally, we show that under standard lipolytic stimulation, 3T3-L1 adipocytes do not release cholesterol onto apoA-I, a process that required long exposures to lipolytic agents (24 h). In conclusion, despite large induction of ABCA1 mRNA during differentiation, cholesterol efflux through the ABCA1 pathway remains limited in adipocytes and requires prolonged lipolysis. This is consistent with the view of the adipocyte behaving as a cholesterol sink, with plasma cholesterol-buffering properties.  (+info)

(4/1584) Activation of PPARalpha and PPARgamma by environmental phthalate monoesters.

Phthalate esters are widely used as plasticizers in the manufacture of products made of polyvinyl chloride. Mono-(2-ethylhexyl)-phthalate (MEHP) induces rodent hepatocarcinogenesis by a mechanism that involves activation of the nuclear transcription factor peroxisome proliferator-activated receptor-alpha (PPARalpha). MEHP also activates PPAR-gamma (PPARgamma), which contributes to adipocyte differentiation and insulin sensitization. Human exposure to other phthalate monoesters, including metabolites of di-n-butyl phthalate and butyl benzyl phthalate, is substantially higher than that of MEHP, prompting this investigation of their potential for PPAR activation, assayed in COS cells and in PPAR-responsive liver (PPARalpha) and adipocyte (PPARgamma) cell lines. Monobenzyl phthalate (MBzP) and mono-sec-butyl phthalate (MBuP) both increased the COS cell transcriptional activity of mouse PPARalpha, with effective concentration for half-maximal response (EC50) values of 21 and 63 microM, respectively. MBzP also activated human PPARalpha (EC50=30 microM) and mouse and human PPARgamma (EC50=75-100 microM). MEHP was a more potent PPAR activator than MBzP or MBuP, with mouse PPARalpha more sensitive to MEHP (EC50=0.6 microM) than human PPARalpha (EC50=3.2 microM). MEHP activation of PPARgamma required somewhat higher concentrations, EC50=10.1 microM (mouse PPARgamma) and 6.2 microM (human PPARgamma). No significant PPAR activation was observed with the monomethyl, mono-n-butyl, dimethyl, or diethyl esters of phthalic acid. PPARalpha activation was verified in FAO rat liver cells stably transfected with PPARalpha, where expression of several endogenous PPARalpha target genes was induced by MBzP, MBuP, and MEHP. Similarly, activation of endogenous PPARgamma target genes was evidenced for all three phthalates by the stimulation of PPARgamma-dependent adipogenesis in the 3T3-L1 cell differentiation model. These findings demonstrate the potential of environmental phthalate monoesters for activation of rodent and human PPARs and may help to elucidate the molecular basis for the adverse health effects proposed to be associated with human phthalate exposure.  (+info)

(5/1584) Mutational analysis of the hormone-sensitive lipase translocation reaction in adipocytes.

Lipolysis in adipocytes governs the release of fatty acids for the supply of energy to various tissues of the body. This reaction is mediated by hormone-sensitive lipase (HSL), a cytosolic enzyme, and perilipin, which coats the lipid droplet surface in adipocytes. Both HSL and perilipin are substrates for polyphosphorylation by protein kinase A (PKA), and phosphorylation of perilipin is required to induce HSL to translocate from the cytosol to the surface of the lipid droplet, a critical step in the lipolytic reaction (Sztalryd C., Xu, G., Dorward, H., Tansey, J. T., Contreras, J.A, Kimmel, A. R., and Londos, C. (2003) J. Cell Biol. 161, 1093-1103). In the present paper we demonstrate that phosphorylation at one of the two more recently discovered PKA sites within HSL, serines 659 and 660, is also required to effect the translocation reaction. Translocation does not occur when these serines residues are mutated simultaneously to alanines. Also, mutation of the catalytic Ser-423 eliminates HSL translocation, showing that the inactive enzyme does not migrate to the lipid droplet upon PKA activation. Thus, HSL translocation requires the phosphorylation of both HSL and perilipin.  (+info)

(6/1584) Insulin stimulates expression of the pyruvate kinase M gene in 3T3-L1 adipocytes.

M2-type pyruvate kinase (M2-PK) mRNA is produced from the PKM gene by an alternative RNA splicing in adipocytes. We found that insulin increased the level of M2-PK mRNA in 3T3-L1 adipocytes in both time- and dose-dependent manners. This induction did not require the presence of glucose or glucosamine in the medium. The insulin effect was blocked by pharmacological inhibitors of insulin signaling pathways such as wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI3K), and PD98059, an inhibitor of mitogen-activated protein kinase (MAPK) kinase. A stable reporter expression assay showed that the promoter activity of an about 2.2-kb 5'-flanking region of the rat PKM gene was stimulated by insulin, but the extents of these stimulations were lower than those of the mRNA stimulation. Thus, we suggest that insulin increases the level of M2-PK mRNA in adipocytes by acting at transcriptional and post-transcriptional levels through signaling pathways involving both PI3K and MAPK kinase.  (+info)

(7/1584) Syntaxin 6 regulates Glut4 trafficking in 3T3-L1 adipocytes.

Insulin stimulates the movement of glucose transporter-4 (Glut4)-containing vesicles to the plasma membrane of adipose cells. We investigated the role of post-Golgi t-soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) in the trafficking of Glut4 in 3T3-L1 adipocytes. Greater than 85% of syntaxin 6 was found in Glut4-containing vesicles, and this t-SNARE exhibited insulin-stimulated movement to the plasma membrane. In contrast, the colocalization of Glut4 with syntaxin 7, 8, or 12/13 was limited and these molecules did not translocate to the plasma membrane. We used adenovirus to overexpress the cytosolic domain of these syntaxin's and studied their effects on Glut4 traffic. Overexpression of the cytosolic domain of syntaxin 6 did not affect insulin-stimulated glucose transport, but increased basal deGlc transport and cell surface Glut4 levels. Moreover, the syntaxin 6 cytosolic domain significantly reduced the rate of Glut4 reinternalization after insulin withdrawal and perturbed subendosomal Glut4 sorting; the corresponding domains of syntaxins 8 and 12 were without effect. Our data suggest that syntaxin 6 is involved in a membrane-trafficking step that sequesters Glut4 away from traffic destined for the plasma membrane. We speculate that this is at the level of traffic of Glut4 into its unique storage compartment and that syntaxin 16 may be involved.  (+info)

(8/1584) HDL-mediated cholesterol uptake and targeting to lipid droplets in adipocytes.

Adipocytes express high levels of the HDL scavenger receptor class B type I in a differentiation-dependent manner. We thus have analyzed the routes of HDL cholesterol trafficking at different phases of adipocyte differentiation in the 3T3-L1 cell line. One novel and salient feature of this paper is the observation of a widespread distribution in the cell cytoplasm of Golgi markers, caveolin-2, and a fluorescent cholesterol analog NBD-cholesterol (NBD-chol), observed in the early phases of adipocyte formation, clearly distinct from that observed in mature fat cells (i.e., with fully formed lipid vesicles). Thus, in cells without visible lipid droplets, Golgi markers (Golgi 58K, Golgin 97, trans-Golgi network 38, Rab 6, and BODIPY-ceramide), caveolin-2, and NBD-chol all colocalize in a widespread distribution in the cell. In contrast, when lipid droplets are fully formed at latter stages, these markers clearly are distributed to distinct cell compartments: a compact juxtanuclear structure for the Golgi markers and caveolin-2, while NDB-chol concentrates in lipid droplets. In addition, disorganization of the Golgi using three different agents (Brefeldin, monensin, and N-ethyl-maleimide) drastically reduces NBD-chol uptake at different phases of adipocyte formation, strongly suggesting that the Golgi apparatus plays a critical role in HDL-mediated NBD uptake and routing to lipid droplets.  (+info)


  • Downstream events involve the activation of protein kinase B (Akt), protein kinase A (PKA), and WNK1 ( 5 ), leading ultimately to branching intracellular pathways, inducing glucose uptake and regulating cell metabolism, growth, and differentiation. (

glucose uptake

  • Furthermore, NOD1 activation suppresses insulin signaling, as revealed by attenuated tyrosine phosphorylation and increased inhibitory serine phosphorylation, of IRS-1 and attenuated phosphorylation of Akt and downstream target GSK3α/3β, resulting in decreased insulin-induced glucose uptake in 3T3-L1 adipocytes. (


  • NOD1 and NOD2 mRNA are markedly increased in differentiated murine 3T3-L1 adipocytes and human primary adipocyte culture upon adipocyte conversion. (
  • Stimulation of NOD1 with a synthetic ligand Tri-DAP induces proinflammatory chemokine MCP-1, RANTES, and cytokine TNF-α and MIP-2 (human IL-8 homolog) and IL-6 mRNA expression in 3T3-L1 adipocytes in a time- and dose-dependent manner. (
  • These studies were followed by the investigations of TLRs in 3T3-L1 adipocytes and in vivo in TLR4 or TLR2 knockout mice. (
  • Consistent with these results, a broad mRNA expression profile of TLRs and their functionalities have been demonstrated in mature 3T3-L1 adipocytes ( 25 ) and human primary adipocytes isolated from fat tissue ( 24 ). (
  • Niacin had no effect on adiponectin secretion or lipolysis in 3T3-L1 adipocytes, which have limited cell surface expression of the GPR109A receptor. (
  • GPR109A (PUMA-G in mice and HM74A in humans) is a seven-transmembrane G protein-coupled receptor of the G i family that is expressed mainly in white adipocytes and immune cells such as monocytes and neutrophils ( 23 ). (
  • Western blot analysis of extracts from 3T3-L1 cells and human adipocytes using SIK2 (D28G3) Rabbit mAb. (


  • Active Motif offers high-quality nuclear, cytoplasmic and whole-cell extracts from a variety of cell types and tissue sources, many prepared from cells that were treated to specifically induce activation of hard-to-detect transcription factors. (


  • We and others have shown that TLR4 and TLR2 (dimerized with TLR6 or TLR1) can mediate SFA-induced NF-κB activation and target gene response in murine macrophage RAW264.7 cells ( 26 , 28 ) and myotube C 2 C 12 cells ( 35 ), respectively. (
  • Oxidized Phospholipids Stimulate Tissue Factor Expression in Human Endothelial Cells Via Activation of ERK/EGR-1 and Ca(++)/NFAT Blood. (
  • Pubmed ID: 11756172 Activation of endothelial cells by lipid oxidation products is a key event in the initiation and progression of the atherosclerotic lesion. (

gene expression

  • To our knowledge, this is the first study delivering an in depth mechanistic outline of the mode of action of TBT as an obesogen, combining effects on both cell physiological and gene expression level. (
  • Validated MISSION ® siRNAs have been functionally validated in Hela cells by Sigma scientists to Knockdown gene expression by 75% or greater. (


  • Minimally modified low-density lipoprotein (MM-LDL) induces the expression of certain inflammatory molecules such as tissue factor (TF) in endothelial cells. (


  • Glycerol Production from Glucose and Fructose by 3T3-L1 Cells: A Mechanism of Adipocyte Defense from Excess Substrate. (
  • Incubation of hepatocyte, adipocyte and β-cell lines with cholic acid decreased ER stress. (


  • Omega-3 fatty acids decreased by nearly one half relative to omega-6 fatty acids in PTB knockdown cells compared with controls, with a particularly strong decrease in eicosapentaenoic acid (EPA) concentration and its ratio to arachidonic acid (ARA). (


  • In HepG2 cells, PTB knockdown modulated alternative splicing of FADS2, as well as FADS3, a putative desaturase of unknown function. (


  • OxPAPC induced TF activity and protein expression in human umbilical vein endothelial cells (HUVECs). (


  • Moreover, mice with TLR2 deficiency were also protected from high-fat diet-induced insulin resistance, tissue inflammation, and pancreatic β-cell dysfunction ( 6 ). (
  • Amelioration of ER stress coincided with improved insulin signaling and preservation of β-cell mass in IT-operated animals. (
  • We now report that mammalian neuraminidase 1 (Neu1), in addition to its catabolic function in lysosomes, is transported to the cell surface where it is involved in the regulation of insulin signaling. (
  • Cells from sialidosis patients with a genetic deficiency of Neu1 show impairment of insulin-induced phosphorylation of downstream protein kinase AKT, and treatment of these cells with purified Neu1 restores signaling. (
  • The signaling cascade starts from binding of insulin to the cell surface insulin receptor kinase (IRK). (
  • At the same time, a pool of inactive, nonphosphorylated IRK, present in the cell membrane and in endosomes, can be substantial, especially in vitro at saturating insulin concentrations ( 11 ). (


  • If you can't find the extract you're looking for, our Nuclear Extract Kit makes it easy to consistently isolate high yields of nuclear, cytoplasmic or whole-cell extract from mammalian cell or tissue samples. (
  • Cell Tissue Res 342:1-11. (


  • These findings reveal a novel role for PTB, regulating availability of membrane components and eicosanoid precursors for cell signaling. (


  • This study was designed to unravel the molecular mechanisms of the model obesogen TBT, using microarray analysis in the 3T3-L1 in vitro system, and to evaluate the use of toxicogenomics for obesogen screening. (


  • Seminars in Cell & Developmental Biology , Vol. 23, Issue. (


  • in that case my transfection is never gonna be stable because over a certain period of time my cells can decide that they just wanna keep the AB resistance and throw the other one out and i wouldnt notice it. (
  • Treat cells by adding fresh media containing regulator for desired time. (


  • Cell culture models were used to investigate interactions between bile acids and ER stress. (


  • Furthermore, our results show that combining transcriptomics with 3T3-L1 cells is a promising tool for screening of potential obesogenic compounds. (


  • Select the wells with only one island of cells and grow them further in bigger wells to screen for eGFP positivity and the other protein which is expressed. (


  • formation on the level of individual endothelial cells was visualized by combining geNOps with a chemical Ca 2+ sensor. (


  • where GFP- At RabF2b was transiently expressed in cell lines in which the ARF-GEF, GNOM, was mutated. (


  • In this study, we investigate the location and function of a member of this family, At RabF2b (Ara7) in tobacco ( Nicotiana tabacum ) leaf epidermal cells using a live cell imaging approach. (


  • 2 weeks and dilute them in a 96 well plate untill you have about 1 cells per well (at first not visible). (
  • Lyse cells by adding 1X SDS sample buffer (100 µl per well of 6-well plate or 500 µl for a 10 cm diameter plate). (
  • Immediately scrape the cells off the plate and transfer the extract to a microcentrifuge tube. (


  • Sonicate for 10-15 sec to complete cell lysis and shear DNA (to reduce sample viscosity). (