2 edition of Morphological characterization of perinuclear GLUT4 distribution in L6 myoblasts. found in the catalog.
Morphological characterization of perinuclear GLUT4 distribution in L6 myoblasts.
Chandrasagar B. Dugani
Written in English
Glucose transporter-4 (GLUT4) cycles to and from the plasma membrane and shows predominant perinuclear localization in unstimulated muscle and adipose tissues. This area is also occupied by recycling endosomes (RE), the Golgi complex, and the trans-Golgi network. Thorough characterization of insulin-responsive perinuclear compartments is lacking. Therefore, we evaluated insulin"s effect on (i) the presence of GLUT4 in these compartments and (ii) the dynamics of perinuclear GLUT4 redistribution. We observed that insulin stimulation reduces the association of GLUT4 only with the RE and induces perinuclear GLUT4 remodelling that parallels the exocytic and internalization profiles of the transporter. Insulin-mediated GLUT4 translocation requires input from phosphatidylinositol-3-kinase (PI3-K), but the steps regulated are unknown. We show that mutants of (PI3-K), and its downstream molecule Akt Substrate 160 kDa (AS160) inhibit insulin-induced perinuclear GLUT4 remodelling. In summary, we characterize a novel insulin effect of perinuclear GLUT4 remodelling and demonstrate that it is regulated by the (PI3-K), → AS160 pathway.
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Differences between the genomes of lymphoblastoid cell lines and blood-derived samples Lena M Joesch-Cohen, Gustavo Glusman Institute for Systems Biology, Seattle, WA, USA Abstract: Lymphoblastoid cell lines (LCLs) represent a convenient research tool for expanding the amount of biologic material available from an individual. LCLs are commonly used as reference materials, most Cited by: 3. 3. Definitions. Purified Water - water from a deionizing system, resistivity > or = 18MΩ•cm @ 25ºC. Unit Definition - One unit will oxidize μmole of D-glucosephosphate to 6-phospho-D-gluconate per minute in the presence of β-NADP at pH at 25°C.
Expression of POGLUT1 (K23Rik, C3orf9, hCLP46, KDELCL1, KTELC1, MDS, MDSRP, MGC, Rumi) in placenta tissue. Antibody staining with HPA and. Precise Probing of Residue Roles by Post-Translational β,γ-C,N Aza- Michael Mutagenesis in Enzyme Active Sites Jitka Dadová, † Kuan-Jung Wu, † Patrick G. Isenegger, † .
RNA tissue distribution i The RNA distribution category is based on mRNA expression levels in the analyzed samples based on a combination of data from HPA, GTEX and FANTOM5. The categories include: detected in all, detected in many, detected in some, detected in single and not detected. Research articleGene expression results in inCited by:
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Plasmids, cells, and transfections. GLUT4myc cDNA was constructed by inserting the human c-myc epitope (14 amino acids) into the first ectodomain of GLUT4, subcloned into the pCXN2 vector, and stably transfected into L6 myoblasts (26, 29).The open reading frame of the bovine PKB cDNA was cloned into the eukaryotic expression vector pcDNA3 (Invitrogen) and in the process was fused to an N Cited by: (A) Schematic presentation of confocal image analysis of GLUT4-myc translocation.
(B) Comparison of GLUT4 translocation in L6 myoblasts, L6-GLUT4-myc cells, and L6-AS_v2-GLUT4 cells after stimulation with nM insulin. L6-AS_v2-GLUT4 cells were incubated with doxycycline to induce expression of AS_v2 protein or were left by: 8.
In contrast, the activation of all three Akt isoforms and GLUT4 translocation are similar in myoblasts and myotubes. Furthermore, GLUT4myc translocation and phosphorylation of regulatory sites on Akt in L6-GLUT4myc myotubes are equally sensitive to insulin, whereas glucose uptake and phosphorylation of regulatory sites on p38 MAPK show lower.
In myoblasts isolated from a healthy individual (no insulin resistance), insulin induces a dose-dependent translocation of GLUT4 from the cytoplasm to the plasma membrane (Figure 1, left).
In contrast, insulin does not induce GLUT4 translocation to the plasma membrane of myoblasts isolated from a patient with insulin resistance (Figure 1, right).Cited by: The use of L6 cells overexpressing AS_v2 can be considered as a novel tool for the characterization of molecules modulating insulin signaling and GLUT4 translocation, and an image-based assay.
For GLUT4 vesicle fusion to occur, GLUT4 vesicles must first be tethered to the plasma membrane. A key tethering factor in exocytosis is a hetero-octameric protein complex called the exocyst. The parental cell lines L6-wt and L6-GLUT4-myc were used as controls. In L6-wt cells, that did not express GLUT4-myc, no translocation was measured neither under basal nor insulin-stimulated conditions.
In L6-GLUT4-myc cells a slight increase of GLUT4 translocation was observed after exposure to nM by: Acknowledgments. The authors would like to thank Professor Y.
Ebina for help in the development of the stable L6-GLUT4myc cell line and Dr. Bilan for the careful reading of this manuscript. This work was supported by a Canadian Institutes of Health Research (CIHR) grant (MT) to AK. C.N.A. has received doctoral studentship funding from an Ontario Graduate Scholarship (OGS) and a Doctoral Cited by: Using L6 rat skeletal myoblasts stably expressing myc-tagged GLUT4, we found that Arp2/3, acting downstream of Rac GTPase, is responsible for the cortical actin polymerization evoked by insulin.
siRNA-mediated silencing of either Arp3 or p34 subunits of the Arp2/3 complex abrogated actin remodeling and impaired GLUT4 translocation. Characterization of p Localization and Function by Immunofluorescence. As shown in Figure 5A, p colocalizes with GLUT4 in the perinuclear region of 3T3-L1 adipocytes under basal conditions (panels a–c).
Unlike GLUT4, the distribution of. 1. Biochem Biophys Res Commun. Dec 1;67(3) Evidence that a membrane bound lectin mediates fusion of L6 myoblasts.
Gartner TK, Podleski by: Western University [email protected] Digitized Theses Digitized Special Collections Cell Surface Phosphorylation And Myogenesis In L6 Rat Myoblasts. A new generation of polyimide (PI)/silver nanowires nanocomposites were prepared by a solution mixing method with different contents of prepared silver nanowires.
The structural and properties of pure PI and PI/Ag nanocomposites were studied by several techniques such as fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance spectroscopy, field emission scanning Cited by: 8.
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Start studying Endo III. Learn vocabulary, terms, and more with flashcards, games, and other study tools. Start studying ABI Chapter 9 Review Questions (Select).
Learn vocabulary, terms, and more with flashcards, games, and other study tools. I'm relatively new to cell culture and I'm having problems inducing my L6's to differentiate to myotubes.
Here's the details: I'm using 10mm dishes with 10mL alpha-MEM, 2% fetal calf serum and 1% penn/strep. Differentiation media added when cells were roughly 80% confluent.
Cells are passage Part of the ‘acyl-lipid metabolism’ chapter in The Arabidopsis Book Page 1 4. SUMMARY OF ARABIDOPSIS LIPID COMPOSITION This section summarizes the acyl lipid composition for various tissues and organs of wild type Arabidopsis.
Data presented in this chapter were collected from Col-0 ecotype unless otherwise Size: KB. Immunofluorescence of Plasma Membrane Sheets. Translocation of GLUT4 to the cell surface was assessed using an immunofluorescence staining of plasma membrane sheets ().
3T3-L1 cells were grown and differentiated on glass ing the indicated treatments, the coverslips were washed in ice-cold buffer containing 50 mM Hepes (pH ) and mM NaCl. Every Step of the Way, a Wide Range of Cell Health Products.
Maintaining healthy cells is the key to experimental success and reproducible research results. Three phospholipase Ds (PLDs; EC ) have been cloned from Arabidopsis, and they exhibit two distinct types of activities: polyphosphoinositide-requiring PLDβ and PLDγ, and polyphosphoinositide-independent PLDα.
In subcellular fractions of Arabidopsis leaves, PLDα and PLDγ were both present in the plasma membrane, intracellular membranes, mitochondria, and clathrin .Zheng et al. find that, in mESCs without any lamins, TAD structures are not affected, but TAD-TAD interactions are changed.
Different lamina-associated chromatin domains (LADs) are decondensed or detached from the nuclear periphery to gain contact with non-LADs.
This leads to global transcriptional changes not limited to genes in by: o GLUT 1 – normal affinity glucose. Found in most cells, including erythrocytes o GLUT 2 – low affinity for glucose (i.e. glucose must be present in high concentration in order for transport to happen.
Found in proximal tubule of kidneys, β-cells of pancreas, intestinal epithelium, and hepatocytes. o GLUT 3 – highest affinity for glucose. Found in neurons.