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eukaryotic translation initiation factor 4e binding protein  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc eukaryotic translation initiation factor 4e binding protein
    Resistance exercise exceeds the benefits of endurance exercise in ameliorating metabolic dysfunction. Following 8 weeks of diet and exercise interventions, all mice were assessed for their metabolic function by GTT, ITT, and skeletal muscle response of Akt and AS160 phosphorylation to injection of insulin measured by Western blot. (A–C) HOMA-IR taken after an overnight fast for baseline glucose and insulin. (D and E) GTT from 0–120 min and calculated AUC; colored * indicates significant difference from NC-SED. (F and G) ITT from 0–60 min and calculated AUC; colored * indicates significant difference from NC-SED. (H–N) pAkt stimulation, AS160 S318, and AS160 T642 in hindlimb muscles before and after insulin injection and the pre–post ∆ in phosphorylation. (O and P) Total and phosphorylated <t>4E-BP1.</t> (Q–T) Western results for Raptor, COX4, LC3 II/I, and ubiquitin staining. Representative western blot images inset right. Data presented as mean ± standard error of the mean. Statistical analysis performed by analysis of variance between groups: * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. NC-SED n : 8–16 (white); HFD-SED n : 8–18 (red); HFD-R EX n : 8–16 (blue); HFD-E EX n : 8–15 (green). 4E-BP1 = <t>Eukaryotic</t> translation initiation factor 4E binding protein; Akt = protein kinase B; AS160 = Akt substrate 160; COX4 = cytochrome c oxidase 4; CS = citrate sythase; E EX = endurance exercise; GAPDH = glyceraldehyde 3 phosphate dehydrogenase; GTT = glucose tolerance test; HFD = high fat diet; HOMA-IR = homeostatic model assessment for insulin resistance; iAUC = integrated area under the curve; ITT = insulin tolerance test; LC3 II/I = microtubule-associated protein light chain 3; NC = normal chow; pAkt = phospho-Akt; R EX = resistance exercise; SED = sedentary; Ub = ubiquitin.
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    Images

    1) Product Images from "Weightlifting outperforms voluntary wheel running for improving adiposity and insulin sensitivity in obese mice"

    Article Title: Weightlifting outperforms voluntary wheel running for improving adiposity and insulin sensitivity in obese mice

    Journal: Journal of Sport and Health Science

    doi: 10.1016/j.jshs.2025.101100

    Resistance exercise exceeds the benefits of endurance exercise in ameliorating metabolic dysfunction. Following 8 weeks of diet and exercise interventions, all mice were assessed for their metabolic function by GTT, ITT, and skeletal muscle response of Akt and AS160 phosphorylation to injection of insulin measured by Western blot. (A–C) HOMA-IR taken after an overnight fast for baseline glucose and insulin. (D and E) GTT from 0–120 min and calculated AUC; colored * indicates significant difference from NC-SED. (F and G) ITT from 0–60 min and calculated AUC; colored * indicates significant difference from NC-SED. (H–N) pAkt stimulation, AS160 S318, and AS160 T642 in hindlimb muscles before and after insulin injection and the pre–post ∆ in phosphorylation. (O and P) Total and phosphorylated 4E-BP1. (Q–T) Western results for Raptor, COX4, LC3 II/I, and ubiquitin staining. Representative western blot images inset right. Data presented as mean ± standard error of the mean. Statistical analysis performed by analysis of variance between groups: * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. NC-SED n : 8–16 (white); HFD-SED n : 8–18 (red); HFD-R EX n : 8–16 (blue); HFD-E EX n : 8–15 (green). 4E-BP1 = Eukaryotic translation initiation factor 4E binding protein; Akt = protein kinase B; AS160 = Akt substrate 160; COX4 = cytochrome c oxidase 4; CS = citrate sythase; E EX = endurance exercise; GAPDH = glyceraldehyde 3 phosphate dehydrogenase; GTT = glucose tolerance test; HFD = high fat diet; HOMA-IR = homeostatic model assessment for insulin resistance; iAUC = integrated area under the curve; ITT = insulin tolerance test; LC3 II/I = microtubule-associated protein light chain 3; NC = normal chow; pAkt = phospho-Akt; R EX = resistance exercise; SED = sedentary; Ub = ubiquitin.
    Figure Legend Snippet: Resistance exercise exceeds the benefits of endurance exercise in ameliorating metabolic dysfunction. Following 8 weeks of diet and exercise interventions, all mice were assessed for their metabolic function by GTT, ITT, and skeletal muscle response of Akt and AS160 phosphorylation to injection of insulin measured by Western blot. (A–C) HOMA-IR taken after an overnight fast for baseline glucose and insulin. (D and E) GTT from 0–120 min and calculated AUC; colored * indicates significant difference from NC-SED. (F and G) ITT from 0–60 min and calculated AUC; colored * indicates significant difference from NC-SED. (H–N) pAkt stimulation, AS160 S318, and AS160 T642 in hindlimb muscles before and after insulin injection and the pre–post ∆ in phosphorylation. (O and P) Total and phosphorylated 4E-BP1. (Q–T) Western results for Raptor, COX4, LC3 II/I, and ubiquitin staining. Representative western blot images inset right. Data presented as mean ± standard error of the mean. Statistical analysis performed by analysis of variance between groups: * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. NC-SED n : 8–16 (white); HFD-SED n : 8–18 (red); HFD-R EX n : 8–16 (blue); HFD-E EX n : 8–15 (green). 4E-BP1 = Eukaryotic translation initiation factor 4E binding protein; Akt = protein kinase B; AS160 = Akt substrate 160; COX4 = cytochrome c oxidase 4; CS = citrate sythase; E EX = endurance exercise; GAPDH = glyceraldehyde 3 phosphate dehydrogenase; GTT = glucose tolerance test; HFD = high fat diet; HOMA-IR = homeostatic model assessment for insulin resistance; iAUC = integrated area under the curve; ITT = insulin tolerance test; LC3 II/I = microtubule-associated protein light chain 3; NC = normal chow; pAkt = phospho-Akt; R EX = resistance exercise; SED = sedentary; Ub = ubiquitin.

    Techniques Used: Phospho-proteomics, Injection, Western Blot, Muscles, Ubiquitin Proteomics, Staining, Binding Assay

    Related Articles

    Ubiquitin Proteomics:

    Article Title: Low dose radiation induces new protein synthesis that promotes cancer radiotherapy resistance.
    Article Snippet: Radiotherapy (RT) is a cornerstone of cancer treatment; however, its efficacy is frequently hampered by its adverse effects on normal tissues.. By studying the effects of high-dose radiotherapy (HDRT) and low-dose radiotherapy (LDRT), we found that cancer cells adapt distinct responses to these doses to reduce cytotoxicity.. Upon HDRT, cancThese authors contributed equally to this worker cells initiate a strong DNA damage response (DDR) to gain resistance through rapid production and/or activation of proteins for cell cycle arrest and DNA damage repair.

    Blocking Assay:

    Article Title: Salvianolic acid B attenuates cellular senescence and age-related decline in muscle function via dual mTOR/TP53INP2-autophagy regulation
    Article Snippet: Electrophoresis was conducted at 80 V for 30 min followed by 120 V for 90 min. Proteins were transferred to PVDF membranes (Millipore) via wet transfer in Tris-glycine buffer with 20% methanol at 90 V (60 min) for low-molecular-weight proteins or 100 V (90 min) for larger proteins. .. After blocking with 5% BSA for 1 h, membranes were incubated with primary antibodies against LC3B (1:1,000, Proteintech), p62 (1:1,000, MCE), β-actin (1:200,000, Abclonal), p-mTOR(1:1,000,CST), p-S6K(1:1,000, CST), 4E-BP1 (1:1,000, CST) and GAPDH (1:200,000, Abclonal) at 4 °C overnight, followed by HRP-conjugated secondary antibody (1:2000, CST) incubation for 2 h at 37 °C. .. Signals were detected by ECL (Millipore) and quantified using ImageJ after imaging with a ChemiDoc XRS + system (Bio-Rad).

    Article Title: Peptide hydrogel boosts the cytotoxic and metabolic fitness of Vγ9Vδ2 T cells in melanoma immunotherapy
    Article Snippet: Equal protein samples were separated by SDS-PAGE and transferred onto PVDF membranes (Millipore, Germany). .. After blocking with 5% BSA, membranes were incubated with primary antibodies against PI3K, AKT, p-AKT, mTOR, p-mTOR, 4E-BP1, and S6K (Cell Signaling Technology, USA). .. After washing, membranes were incubated with HRP-conjugated secondary antibodies, and signals were visualized with enhanced chemiluminescence (Bio-Rad,USA).

    Incubation:

    Article Title: Salvianolic acid B attenuates cellular senescence and age-related decline in muscle function via dual mTOR/TP53INP2-autophagy regulation
    Article Snippet: Electrophoresis was conducted at 80 V for 30 min followed by 120 V for 90 min. Proteins were transferred to PVDF membranes (Millipore) via wet transfer in Tris-glycine buffer with 20% methanol at 90 V (60 min) for low-molecular-weight proteins or 100 V (90 min) for larger proteins. .. After blocking with 5% BSA for 1 h, membranes were incubated with primary antibodies against LC3B (1:1,000, Proteintech), p62 (1:1,000, MCE), β-actin (1:200,000, Abclonal), p-mTOR(1:1,000,CST), p-S6K(1:1,000, CST), 4E-BP1 (1:1,000, CST) and GAPDH (1:200,000, Abclonal) at 4 °C overnight, followed by HRP-conjugated secondary antibody (1:2000, CST) incubation for 2 h at 37 °C. .. Signals were detected by ECL (Millipore) and quantified using ImageJ after imaging with a ChemiDoc XRS + system (Bio-Rad).

    Article Title: Peptide hydrogel boosts the cytotoxic and metabolic fitness of Vγ9Vδ2 T cells in melanoma immunotherapy
    Article Snippet: Equal protein samples were separated by SDS-PAGE and transferred onto PVDF membranes (Millipore, Germany). .. After blocking with 5% BSA, membranes were incubated with primary antibodies against PI3K, AKT, p-AKT, mTOR, p-mTOR, 4E-BP1, and S6K (Cell Signaling Technology, USA). .. After washing, membranes were incubated with HRP-conjugated secondary antibodies, and signals were visualized with enhanced chemiluminescence (Bio-Rad,USA).

    other:

    Article Title: Neurons with granulovacuolar degeneration bodies are resilient to tau-induced protein synthesis impairment
    Article Snippet: 4E-BP1 , , Rabbit monoclonal , , 1:1000 , Cell Signaling Technology , 9644.

    Article Title: Effects of branched-chain amino acids on iron deficiency-induced muscle atrophy
    Article Snippet: 4E-BP1 , Rabbit , 9452 , 1:1000 , Cell Signaling Technology.

    Article Title: Ad-VT oncolytic adenovirus suppresses bladder cancer via cAMP-dependent AMPK-Raptor activation and G2/M arrest
    Article Snippet: The primary antibodies, CyclinB1 (#4131), p-cdc2 (#4539), cdc2 (#9116), p21 (#2947), AMPK (#5831), p-AMPK (#2535), Raptor (#2280), p-Raptor (#2083), mTOR (#2983), p-mTOR (#5536), S6 (#2317), p-S6 (#4858), 4E-BP1 (#9644), p-4E-BP-1 (#2855), and GAPDH (#5175) were purchased from Cell Signaling Technology.

    Pyrolysis Gas Chromatography:

    Article Title: Limosilactobacillus reuteri ATG-F4 Ameliorates Dexamethasone-Induced Muscle Atrophy through Modulation of Gut Microbiota.
    Article Snippet: Membranes were blocked overnight at 4°C in SuperBlock (PBS) blocking buffer (pH 7.4) containing Kathon antimicrobial agent and incubated with primary antibodies (all from Cell Signaling Technology, USA) at a dilution of 1:1000. .. The following primary antibodies were used: myostatin, p-Akt, Akt, p-FOXO3a, FOXO3a, Atrogin-1, MuRF1, p-AMPK, AMPK, PGC-1α, mtTFA, p-p70S6K, p70S6K, p-rpS6, rpS6, p-4E-BP1, 4E-BP1, and β-actin as a loading control (1:1000, Cell Signaling Technology, USA). .. Blots were washed four times with 0.1% Tween-20 in TBS (TBS-T) before incubation for 1 h at approximately 22–25°C with a goat anti-rabbit IgG HRP-conjugated secondary antibody (Bio-Rad) in TBS-T containing 3% BSA (Bovogen, USA).

    Control:

    Article Title: Limosilactobacillus reuteri ATG-F4 Ameliorates Dexamethasone-Induced Muscle Atrophy through Modulation of Gut Microbiota.
    Article Snippet: Membranes were blocked overnight at 4°C in SuperBlock (PBS) blocking buffer (pH 7.4) containing Kathon antimicrobial agent and incubated with primary antibodies (all from Cell Signaling Technology, USA) at a dilution of 1:1000. .. The following primary antibodies were used: myostatin, p-Akt, Akt, p-FOXO3a, FOXO3a, Atrogin-1, MuRF1, p-AMPK, AMPK, PGC-1α, mtTFA, p-p70S6K, p70S6K, p-rpS6, rpS6, p-4E-BP1, 4E-BP1, and β-actin as a loading control (1:1000, Cell Signaling Technology, USA). .. Blots were washed four times with 0.1% Tween-20 in TBS (TBS-T) before incubation for 1 h at approximately 22–25°C with a goat anti-rabbit IgG HRP-conjugated secondary antibody (Bio-Rad) in TBS-T containing 3% BSA (Bovogen, USA).



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    Resistance exercise exceeds the benefits of endurance exercise in ameliorating metabolic dysfunction. Following 8 weeks of diet and exercise interventions, all mice were assessed for their metabolic function by GTT, ITT, and skeletal muscle response of Akt and AS160 phosphorylation to injection of insulin measured by Western blot. (A–C) HOMA-IR taken after an overnight fast for baseline glucose and insulin. (D and E) GTT from 0–120 min and calculated AUC; colored * indicates significant difference from NC-SED. (F and G) ITT from 0–60 min and calculated AUC; colored * indicates significant difference from NC-SED. (H–N) pAkt stimulation, AS160 S318, and AS160 T642 in hindlimb muscles before and after insulin injection and the pre–post ∆ in phosphorylation. (O and P) Total and phosphorylated 4E-BP1. (Q–T) Western results for Raptor, COX4, LC3 II/I, and ubiquitin staining. Representative western blot images inset right. Data presented as mean ± standard error of the mean. Statistical analysis performed by analysis of variance between groups: * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. NC-SED n : 8–16 (white); HFD-SED n : 8–18 (red); HFD-R EX n : 8–16 (blue); HFD-E EX n : 8–15 (green). 4E-BP1 = Eukaryotic translation initiation factor 4E binding protein; Akt = protein kinase B; AS160 = Akt substrate 160; COX4 = cytochrome c oxidase 4; CS = citrate sythase; E EX = endurance exercise; GAPDH = glyceraldehyde 3 phosphate dehydrogenase; GTT = glucose tolerance test; HFD = high fat diet; HOMA-IR = homeostatic model assessment for insulin resistance; iAUC = integrated area under the curve; ITT = insulin tolerance test; LC3 II/I = microtubule-associated protein light chain 3; NC = normal chow; pAkt = phospho-Akt; R EX = resistance exercise; SED = sedentary; Ub = ubiquitin.

    Journal: Journal of Sport and Health Science

    Article Title: Weightlifting outperforms voluntary wheel running for improving adiposity and insulin sensitivity in obese mice

    doi: 10.1016/j.jshs.2025.101100

    Figure Lengend Snippet: Resistance exercise exceeds the benefits of endurance exercise in ameliorating metabolic dysfunction. Following 8 weeks of diet and exercise interventions, all mice were assessed for their metabolic function by GTT, ITT, and skeletal muscle response of Akt and AS160 phosphorylation to injection of insulin measured by Western blot. (A–C) HOMA-IR taken after an overnight fast for baseline glucose and insulin. (D and E) GTT from 0–120 min and calculated AUC; colored * indicates significant difference from NC-SED. (F and G) ITT from 0–60 min and calculated AUC; colored * indicates significant difference from NC-SED. (H–N) pAkt stimulation, AS160 S318, and AS160 T642 in hindlimb muscles before and after insulin injection and the pre–post ∆ in phosphorylation. (O and P) Total and phosphorylated 4E-BP1. (Q–T) Western results for Raptor, COX4, LC3 II/I, and ubiquitin staining. Representative western blot images inset right. Data presented as mean ± standard error of the mean. Statistical analysis performed by analysis of variance between groups: * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. NC-SED n : 8–16 (white); HFD-SED n : 8–18 (red); HFD-R EX n : 8–16 (blue); HFD-E EX n : 8–15 (green). 4E-BP1 = Eukaryotic translation initiation factor 4E binding protein; Akt = protein kinase B; AS160 = Akt substrate 160; COX4 = cytochrome c oxidase 4; CS = citrate sythase; E EX = endurance exercise; GAPDH = glyceraldehyde 3 phosphate dehydrogenase; GTT = glucose tolerance test; HFD = high fat diet; HOMA-IR = homeostatic model assessment for insulin resistance; iAUC = integrated area under the curve; ITT = insulin tolerance test; LC3 II/I = microtubule-associated protein light chain 3; NC = normal chow; pAkt = phospho-Akt; R EX = resistance exercise; SED = sedentary; Ub = ubiquitin.

    Article Snippet: Primary antibodies used for analysis were from Cell Signaling Technologies (Danvers, MA, USA) and diluted 1:1000 unless otherwise stated as follows: protein kinase B (Akt; 1:500; #4691; Cell Signaling Technologies), phospho-Akt (pAkt) S473 (1:500; #9271; Cell Signaling Technologies), Ubiquitin (#3933; Cell Signaling Technologies), microtubule-associated protein light chain 3 (LC3 II/I; #4018; Cell Signaling Technologies), cytochrome c oxidase subunit 4(COX4; #11967; Cell Signaling Technologies), Akt substrate 160 (AS160 S318; #8619; Cell Signaling Technologies), AS160 T642 (#8881; Cell Signaling Technologies), eukaryotic translation initiation factor 4E binding protein (4E-BP1; #9452; Cell Signaling Technologies), and glyceraldehyde 3 phosphate dehydrogenase (GAPDH; #2118; Cell Signaling Technologies).

    Techniques: Phospho-proteomics, Injection, Western Blot, Muscles, Ubiquitin Proteomics, Staining, Binding Assay

    Effect of corn processing on relative mRNA level for mTOR , 4E-BP1 , p70S6K and protein abundances of mTOR, phosphorylated mTOR (p-mTOR), eukaryotic initiation factor 4E-binding protein 1 (4E-BP1), phosphorylated 4E-BP1 (p-4E-BP1), p70 ribosomal protein S6 kinase (p70S6K) and phosphorylated p70S6K (p-p70S6K) in the longissimus thoracis of piglets. Results are presented as means ± standard error of the mean (SEM), n = 4. Data columns with different letters were significantly different ( P ≤ 0.05).

    Journal: Animal Nutrition

    Article Title: Modulating starch digestion kinetics via feed processing: Implications for growth and metabolism in weaned pigs

    doi: 10.1016/j.aninu.2025.08.011

    Figure Lengend Snippet: Effect of corn processing on relative mRNA level for mTOR , 4E-BP1 , p70S6K and protein abundances of mTOR, phosphorylated mTOR (p-mTOR), eukaryotic initiation factor 4E-binding protein 1 (4E-BP1), phosphorylated 4E-BP1 (p-4E-BP1), p70 ribosomal protein S6 kinase (p70S6K) and phosphorylated p70S6K (p-p70S6K) in the longissimus thoracis of piglets. Results are presented as means ± standard error of the mean (SEM), n = 4. Data columns with different letters were significantly different ( P ≤ 0.05).

    Article Snippet: The membranes were blocked at room temperature, followed by incubation at 4 °C overnight with the following primary antibodies: mammalian target of rapamycin (mTOR, catalog No. 2983, Cell Signaling Technology, Danvers, MA, USA), phosphorylated mTOR (p-mTOR, catalog No. 5536, Cell Signaling Technology, Danvers, MA, USA), eukaryotic initiation factor 4E-binding protein 1 (4E-BP1, catalog No. 9644, Cell Signaling Technology, Danvers, MA, USA), phosphorylated 4E-BP1 (p-4E-BP1, catalog No. 2855, Cell Signaling Technology, Danvers, MA, USA), p70 ribosomal protein S6 kinase (p70S6K, catalog No. 2708, Cell Signaling Technology, Danvers, MA, USA), phosphorylated p70S6K (p-p70S6K, catalog No. 9234, Cell Signaling Technology, Danvers, MA, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, catalog No. 5174, Cell Signaling Technology, Danvers, MA, USA).

    Techniques: Binding Assay