primary antibodies against glut4  (Proteintech)

Journal: Nature Metabolism

Article Title: Skeletal muscle from TBC1D4 p.Arg684Ter variant carriers is severely insulin resistant but exhibits normal metabolic responses during exercise

doi: 10.1038/s42255-024-01153-1

Figure Lengend Snippet: a , Graphical representation of the EHC. b , GIR during the insulin clamp. c – e , GIR ( c ), endogenous glucose rate of appearance (RA) ( d ) and glucose rate of disappearance (RD) ( e ) at 120 min of the clamp. f , Plasma FAs during the clamp. g , Average leg glucose uptake during the last 40 min of the clamp. h , Targeted immunoblotting analyses of proteins associated with glucose metabolism, fat metabolism and the mitochondrial electron transport chain in skeletal muscle. i , Volcano plot showing proteome log 2 fold change (FC) (carriers/controls) plotted against –log 10 P value highlighting downregulated proteins TBC1D4, TBC1D1 and GLUT4 (SLC2A4). *Difference between TBC1D4 carriers and controls at the given time point. ‡Different from basal (time 0) within the same group. n = 5 ( b – g , i ). n = 7 ( h ) except for mTOR, p70S6K and ACC ( n = 5 in controls and TBC1D4 carriers) as well as Akt2, AMPKα2, CS, CD36 and FATP4 ( n = 6) in controls and GLUT1 ( n = 6) in TBC1D4 carriers. Data are means ± s.e.m. Data were analysed using a two-tailed paired Student’s t -test ( c – e , g ), a two-tailed non-paired Student’s t -test ( h ) as well as a two-way repeated ANOVA test (two-factor repeated) and two-tailed Student–Newman–Keuls post hoc analyses for multiple comparisons ( b , f ). Graphics in a created using BioRender.com . a.u., arbitrary units.

Article Snippet: For staining, four to six individual fibres were teased out from the fixed fibre bundles with fine forceps and blocked for 2 h in PBS containing 5% goat serum, 1% BSA and 0.04% saponin and then incubated overnight with primary antibody raised against GLUT4 (rabbit polyclonal; no. PA5-23052; lot WJ3403509, Thermo Scientific).

Techniques: Western Blot, Two Tailed Test

Journal: Nature Metabolism

Article Title: Skeletal muscle from TBC1D4 p.Arg684Ter variant carriers is severely insulin resistant but exhibits normal metabolic responses during exercise

doi: 10.1038/s42255-024-01153-1

Figure Lengend Snippet: a , Representative immunoblots for data related to figure panel 2 h. b , c , Distribution of Myosin Heavy Chains (MHC) ( b ) and regulatory metabolic ( c ) proteins in MHC-defined type 1 and type 2 skeletal muscle fiber bundles. d , Representative immunoblots. e , Quantification of GLUT4 imaging by confocal microscopy of isolated skeletal muscle fibers. n = 7 in Controls and n = 6 in TBC1D4 carriers ( b , c ). n = 15 fibers obtained from 3 subjects (4 to 6 fibers/subject) in each group ( e ). Data are means ± SEM. Data were analyzed using a two-tailed non-paired students t-test ( e ) as well as a two-way repeated ANOVA test (one factor repeated) and two-tailed Student-Newman-Keuls post hoc analyses for multiple comparisons ( b , c ). AU, arbitrary units. Scale bar in panel e is 10 µm.

Article Snippet: For staining, four to six individual fibres were teased out from the fixed fibre bundles with fine forceps and blocked for 2 h in PBS containing 5% goat serum, 1% BSA and 0.04% saponin and then incubated overnight with primary antibody raised against GLUT4 (rabbit polyclonal; no. PA5-23052; lot WJ3403509, Thermo Scientific).

Techniques: Western Blot, Imaging, Confocal Microscopy, Isolation, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Insulin-Activated Signaling Pathway and GLUT4 Membrane Translocation in hiPSC-Derived Cardiomyocytes

doi: 10.3390/ijms25158197

Figure Lengend Snippet: Lactate-purified hiPSC-CMs show significant GLUT4 membrane translocation after insulin stimulation. Bar graph and representative blots of ( a , b ) total lysates and ( c , d ) membrane fractions showing higher GLUT4 levels in membrane fractions of insulin-treated cells. GLUT1 shows no variations after insulin stimulation. (Total lysates: CTRL-GLUT4: 100.00; INS-GLUT4: 97.33 ± 14.33, CTRL-GLUT1: 72.18 ± 12.04; INS-GLUT1: 79.51 ± 22.60; Membrane fractions: CTRL-GLUT4: 100.00; INS-GLUT4: 150.34 ± 5.49, CTRL-GLUT1: 57.26 ± 14.42; INS-GLUT1: 61.04 ± 17.60). ( e ) Representative immunostaining showing GLUT4 membrane translocation in insulin-treated cells (white arrows) (40×, scale bar 10 µm), and ( f ) the bar graph summarizing the immunofluorescence experiments analysis in 34 α-actinin-positive cells for both conditions (CTRL: 1.00 ± 0.13; INS: 2.31 ± 0.28). n = 3 independent experiments, * p < 0.05; *** p < 0.001.

Article Snippet: HiPSC-CMs were dissociated after 14 days and were fixed with 4% formaldehyde for 20 min and later stained with, respectively, primary antibodies against α-actinin (#A7732, Sigma Aldrich, dilution 1:400) and GLUT4 (#PA5-23052, Thermo Fisher Scientific, dilution 1:100) diluted in PBS with 0.2% Triton X-100 and 5% donkey serum for 24 h at 4 °C.

Techniques: Purification, Membrane, Translocation Assay, Immunostaining, Immunofluorescence

Journal: Heliyon

Article Title: Naringenin enhances the efficacy of ferroptosis inducers by attenuating aerobic glycolysis by activating the AMPK-PGC1α signalling axis in liver cancer

doi: 10.1016/j.heliyon.2024.e32288

Figure Lengend Snippet: Primer sequences information.

Article Snippet: Specific primary antibodies against GLUT4 (66846-1-Ig), Cyclin D1 (60186-1-Ig), Cyclin E1 (11554-1-AP) were purchased from Proteintech (Wuhan, China); LDHA (YN033) and GLUT1 (YT1928) were purchased from Immunoway (Hunan, China); PGC1α (#2178) was purchased from CST (Massachusetts, USA); β-actin (ab8227), PDK-1 (ab207450), p-AMPKα1/2 (ab133448), AMPKα1/2 (ab207442), CDK6 (ab124821), CDK2 (ab32147) were purchased from Abcam (United Kingdom).

Techniques:

Journal: Heliyon

Article Title: Naringenin enhances the efficacy of ferroptosis inducers by attenuating aerobic glycolysis by activating the AMPK-PGC1α signalling axis in liver cancer

doi: 10.1016/j.heliyon.2024.e32288

Figure Lengend Snippet: Naringenin inhibits aerobic glycolysis in liver cancer cells. HepG2 cells were treated with Nar (0.1 mM/0.2 mM) and/or erastin (5 μM)/RSL3 (0.1 μM). (A) The glucose uptake levels of the cell. (B) Cellular lactate production. (C)the intracellular ATP level of the cells. (D) Western blot was utilized to determine the protein expression levels of GLUT1, GLUT4, LDHA, and PDK1 in HepG2 cells. (E) qRT-PCR was utilized to determine the mRNA expression levels of GLUT1, GLUT4, LDHA, and PDK1 in HepG2 cells. *P < 0.05, **P < 0.01, ***P < 0.001, n = 3 per group.

Article Snippet: Specific primary antibodies against GLUT4 (66846-1-Ig), Cyclin D1 (60186-1-Ig), Cyclin E1 (11554-1-AP) were purchased from Proteintech (Wuhan, China); LDHA (YN033) and GLUT1 (YT1928) were purchased from Immunoway (Hunan, China); PGC1α (#2178) was purchased from CST (Massachusetts, USA); β-actin (ab8227), PDK-1 (ab207450), p-AMPKα1/2 (ab133448), AMPKα1/2 (ab207442), CDK6 (ab124821), CDK2 (ab32147) were purchased from Abcam (United Kingdom).

Techniques: Western Blot, Expressing, Quantitative RT-PCR