mouse monoclonal antibody against glut4  (R&D Systems)

Journal: Journal of Physiology and Biochemistry

Article Title: D609 inhibition of phosphatidylcholine-specific phospholipase C attenuates prolonged insulin stimulation-mediated GLUT4 downregulation in 3T3-L1 adipocytes

doi: 10.1007/s13105-022-00872-x

Figure Lengend Snippet: Prolonged insulin stimulation requires PC-PLC activity for the downregulation of GLUT4. Prior to stimulation with insulin (500 nM) for 4 h, the 3T3-L1 adipocytes were treated with or without the following compounds at indicated concentrations for 30 min: D609 ( a ), GW4869 ( b ), U73122 ( c ), 1-Butanol ( d ), or HC-3 ( e ). GLUT4 were detected by immunoblotting. The upper panels show the representative immunoblots for GLUT4, and the lower panels were used for quantifying the GLUT4. The results shown are the mean ± SD ( n = 3)

Article Snippet: Monoclonal antibodies against GLUT4 (2213S, 1:1000) were purchased from Cell Signaling (Danvers, MA, USA), and α-tubulin mouse monoclonal antibody (66,031–1-Ig, 1:10,000) was procured from Proteintech (Chicago, IL).

Techniques: Activity Assay, Western Blot

Journal: Journal of Physiology and Biochemistry

Article Title: D609 inhibition of phosphatidylcholine-specific phospholipase C attenuates prolonged insulin stimulation-mediated GLUT4 downregulation in 3T3-L1 adipocytes

doi: 10.1007/s13105-022-00872-x

Figure Lengend Snippet: Effect of exogenous PC-PLC and its products (PCho and DG) on GLUT4 concentrations. The 3T3-L1 adipocytes were treated with or without the following at indicated concentrations for 4 h: PC-PLC ( a ), PCho ( b ), and DG ( c ). After incubating, the cells were lysed and processed for immunoblotting. The upper panels show the representative GLUT4 immunoblots, and the lower panels display the GLUT4 quantification. The results shown are the mean ± SD ( n = 3)

Article Snippet: Monoclonal antibodies against GLUT4 (2213S, 1:1000) were purchased from Cell Signaling (Danvers, MA, USA), and α-tubulin mouse monoclonal antibody (66,031–1-Ig, 1:10,000) was procured from Proteintech (Chicago, IL).

Techniques: Western Blot

Journal: Journal of Traditional and Complementary Medicine

Article Title: A novel phenolic formulation for treating hepatic and peripheral insulin resistance by regulating GLUT4-mediated glucose uptake

doi: 10.1016/j.jtcme.2021.08.004

Figure Lengend Snippet: Effect of phenolic combination and anti-diabetic drugs on IDV-pretreated insulin-resistant L6 myotubes. Palmitate-treated L6 myotubes were stimulated for 5 min without or with 100 μM IDV, then incubated with DMSO (0.1 %), phenolic combination (5 μM EGEG + 10 μM RSV + 25 μM FER), metformin (1 mM) or AICAR (2 mM) for 15 min. Data present the 2DG uptake influenced by palmitate pretreatment only (white bars) and palmitate along with IDV treatment (closed bars). Results are the means +SD of five independent experiments. Different superscript letters indicate significant differences at p < 0.05. The GLUT4 dependent effect above basal was calculated for each stimulator. # p < 0.05 compared with GLUT4 effect in cells not treated with IDV.

Article Snippet: Mouse monoclonal antibody against GLUT4 (clone 1F8) was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Incubation

Journal: Journal of Traditional and Complementary Medicine

Article Title: A novel phenolic formulation for treating hepatic and peripheral insulin resistance by regulating GLUT4-mediated glucose uptake

doi: 10.1016/j.jtcme.2021.08.004

Figure Lengend Snippet: Effect of phenolic combination and anti-diabetic drugs on the translocation of GLUT4 to plasma membrane and the related intracellular signaling molecules including phosphorylated AKT ( p -AKT) and ACC ( p -ACC) in insulin-resistant L6 myotubes. Palmitate-treated myotubes were incubated with DMSO (0.1 %), phenolic combination (5 μM EGEG + 10 μM RSV + 25 μM FER), metformin (1 mM) or AICAR (2 mM) for 15 min. The cell lysates were used for preparation of a plasma membrane fraction as described under “Methods”. Protein (50 μg) was resolved by SDS-PAGE and Western blotted for the proteins shown. Detection was by enhanced chemiluminescence, and a representative blot is depicted. Anti-β-actin was used as protein loading control of whole homogenate.

Article Snippet: Mouse monoclonal antibody against GLUT4 (clone 1F8) was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Translocation Assay, Membrane, Incubation, SDS Page, Western Blot

Journal: Molecular Cancer

Article Title: 3-BrPA eliminates human bladder cancer cells with highly oncogenic signatures via engagement of specific death programs and perturbation of multiple signaling and metabolic determinants

doi: 10.1186/s12943-015-0399-9

Figure Lengend Snippet: Critical regulators of glucose homeostasis are detrimentally targeted after bladder cancer cell exposure to 3-BrPA: drug-induced splicing silencing of GLUT4 RNA. ( a - b ) Representative (three independent experiments) Western blotting profiles of whole-cell protein extracts obtained from RT4, T24 and T24-X cells, seeded at ~60 % confluency and exposed to the indicated doses of 3-BrPA for 24 h. The proteins examined were HK2, GAPDH, p-AS160-Thr 642 , AS160, Rab10, Tug ( a ), GLUT1 and GLUT4 ( b ), while Actin was used as molecule of reference. ( c - f ) Gene expression profiles, as evidenced through employment of RT-sqPCR protocols (three independent experiments), using total RNA preparations derived from RT4, T24 and T24-X cells, grown at ~60 % confluency and treated with the indicated doses of 3-BrPA ( c - f ), Taxol and Doxorubicin ( f ) for 24 h (also, see Additional file : Figure S5, Additional file : Figure S6 and Additional file : Figure S7). Besides the oligonucleotide primers able to specifically recognize the GLUT1 , GLUT2 , GLUT3 ( c ) and FasL ( f ) genes, GLUT4 -specific primers were used to amplify several exon-exon (e.g. Ex7 - Ex8 ), exon-intron (e.g. Ex4 - In4/5 ) and intra-intronic (e.g. In2/3 ) segments of the two major RNA splicing variants examined ( http://www.ensembl.org/Homo_sapiens ) ( d - f ) (also, see Additional file : Figure S5 and Additional file : Table S1). GAPDH served as gene of reference. Ex : (single) exon. In : intron (in-between successive exons). Sv001 / Sv004 : GLUT4 RNA splicing variants

Article Snippet: Mouse monoclonal antibody against GLUT4 was obtained from Santa Cruz Biotechnology Inc. (Texas, USA).

Techniques: Western Blot, Gene Expression, Derivative Assay