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mtor total  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc mtor total
    Current concepts of multiple amino acid–sensing input to mTORC1. AA, amino acid; Akt, RAC-α serine/threonine-protein kinase; ARF1, ADP-ribosylation factor 1; BCAA, branched-chain amino acid; CASTOR1, cytosolic arginine sensor for mTORC1 subunit <t>1;</t> <t>GATOR1/2,</t> GAP activity toward RAGs 1/2; LARS, leucine-tRNA-synthase; LAT1/SLC7A5, L-type amino acid transporter 1; mTORC1, mechanistic target of rapamycin complex 1; PI3K, phosphatidylinositol-3-kinase; RAB1A, Ras-related protein Rab-1A; Rag A/B C/D, RAS-related GTP-binding protein A/B C/D; Ragulator, Late endosomal/lysosomal adaptor and MAPK and <t>mTOR</t> activator; Rheb, Ras homolog enriched in brain; SAM, S-adenosyl methionine; SAMTOR, S-adenosylmethionine sensor upstream of mTORC1; SAR1B, secretion-associated Ras-related GTPase 1B; Sestrin1/2, stress response protein 1/2; SLC38A9, solute carrier family 38 member 9; SNAT2, sodium-coupled neutral amino acid transporter 2; TARS2, threonyl-tRNA synthetase 2; TFEB, transcription factor EB; TSC1/2, tuberous sclerosis complex 1/2; V-ATPase, vacuolar H + -ATPase. Created in BioRender. Ramos dos Santos, A.C. (2026) https://BioRender.com/54b947z .
    Mtor Total, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 8569 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/total-mtor+antibody/mTOR+Antibody/pmc12975357-114-49-54
    Average 97 stars, based on 8569 article reviews
    mtor total - by Bioz Stars, 2026-09
    97/100 stars

    Images

    1) Product Images from "Amino Acid Signaling in Skeletal Muscle Is Blunted by Prematurity in a Piglet Model"

    Article Title: Amino Acid Signaling in Skeletal Muscle Is Blunted by Prematurity in a Piglet Model

    Journal: The Journal of Nutrition

    doi: 10.1016/j.tjnut.2025.101303

    Current concepts of multiple amino acid–sensing input to mTORC1. AA, amino acid; Akt, RAC-α serine/threonine-protein kinase; ARF1, ADP-ribosylation factor 1; BCAA, branched-chain amino acid; CASTOR1, cytosolic arginine sensor for mTORC1 subunit 1; GATOR1/2, GAP activity toward RAGs 1/2; LARS, leucine-tRNA-synthase; LAT1/SLC7A5, L-type amino acid transporter 1; mTORC1, mechanistic target of rapamycin complex 1; PI3K, phosphatidylinositol-3-kinase; RAB1A, Ras-related protein Rab-1A; Rag A/B C/D, RAS-related GTP-binding protein A/B C/D; Ragulator, Late endosomal/lysosomal adaptor and MAPK and mTOR activator; Rheb, Ras homolog enriched in brain; SAM, S-adenosyl methionine; SAMTOR, S-adenosylmethionine sensor upstream of mTORC1; SAR1B, secretion-associated Ras-related GTPase 1B; Sestrin1/2, stress response protein 1/2; SLC38A9, solute carrier family 38 member 9; SNAT2, sodium-coupled neutral amino acid transporter 2; TARS2, threonyl-tRNA synthetase 2; TFEB, transcription factor EB; TSC1/2, tuberous sclerosis complex 1/2; V-ATPase, vacuolar H + -ATPase. Created in BioRender. Ramos dos Santos, A.C. (2026) https://BioRender.com/54b947z .
    Figure Legend Snippet: Current concepts of multiple amino acid–sensing input to mTORC1. AA, amino acid; Akt, RAC-α serine/threonine-protein kinase; ARF1, ADP-ribosylation factor 1; BCAA, branched-chain amino acid; CASTOR1, cytosolic arginine sensor for mTORC1 subunit 1; GATOR1/2, GAP activity toward RAGs 1/2; LARS, leucine-tRNA-synthase; LAT1/SLC7A5, L-type amino acid transporter 1; mTORC1, mechanistic target of rapamycin complex 1; PI3K, phosphatidylinositol-3-kinase; RAB1A, Ras-related protein Rab-1A; Rag A/B C/D, RAS-related GTP-binding protein A/B C/D; Ragulator, Late endosomal/lysosomal adaptor and MAPK and mTOR activator; Rheb, Ras homolog enriched in brain; SAM, S-adenosyl methionine; SAMTOR, S-adenosylmethionine sensor upstream of mTORC1; SAR1B, secretion-associated Ras-related GTPase 1B; Sestrin1/2, stress response protein 1/2; SLC38A9, solute carrier family 38 member 9; SNAT2, sodium-coupled neutral amino acid transporter 2; TARS2, threonyl-tRNA synthetase 2; TFEB, transcription factor EB; TSC1/2, tuberous sclerosis complex 1/2; V-ATPase, vacuolar H + -ATPase. Created in BioRender. Ramos dos Santos, A.C. (2026) https://BioRender.com/54b947z .

    Techniques Used: Activity Assay, Binding Assay

    Related Articles

    Saline:

    Article Title: Resistance exercise enhances cognitive function in mouse.
    Article Snippet: ▼ Physical exercise has been shown to increase adult neurogenesis in the hippocampus and to enhance synaptic plasticity.. It has been demonstrated that these neuroprotective eff ects can be observed following aerobic exercise.. However, it remains unknown whether plasticity molecules, such as brain-derived neurotrophic factor (BDNF) and cyclic AMP response element-binding protein (CREB), are expressed in the hippocampus following resistance exercise.

    Article Title: Lower basal and postprandial muscle protein synthesis after 2 weeks single‐leg immobilization in older men: No protective effect of anti‐inflammatory medication
    Article Snippet: The primary antibodies from Cell Signaling Technology, Danvers, MA were total‐mTOR (no. 2983) diluted 1:1000, total‐p70‐S6K1 (no. 2708) diluted 1:1000, p‐p70‐S6K (Thr389; no. 9206) diluted 1:1000, total‐AKT1 (no. 2967) diluted 1:1000, p‐AKT (Thr308; no. 2965) diluted 1:1000, p‐eEF2 (Thr56; no. 2331) diluted 1:2000 and p‐4E‐BP1 (Thr37/46; no. 2855) diluted 1:1000, and from Abcam, Cambridge, MA total‐eEF2 (ab130187) diluted 1:500.

    Article Title: Rapamycin and mTORC2 inhibition synergistically reduce contraction‐stimulated muscle protein synthesis
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    Article Title: Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/mTOR Pathway
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    Article Title: Progranulin depletion inhibits proliferation via the transforming growth factor beta/SMAD family member 2 signaling axis in Kasumi-1 cells
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    Article Title: A CCL2/ROS autoregulation loop is critical for cancer-associated fibroblasts-enhanced tumor growth of oral squamous cell carcinoma.
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    Article Title: Homer Interactions Are Necessary for Metabotropic Glutamate Receptor-Induced Long-Term Depression and Translational Activation
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    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity
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    Incubation:

    Article Title: Resistance exercise enhances cognitive function in mouse.
    Article Snippet: ▼ Physical exercise has been shown to increase adult neurogenesis in the hippocampus and to enhance synaptic plasticity.. It has been demonstrated that these neuroprotective eff ects can be observed following aerobic exercise.. However, it remains unknown whether plasticity molecules, such as brain-derived neurotrophic factor (BDNF) and cyclic AMP response element-binding protein (CREB), are expressed in the hippocampus following resistance exercise.

    Article Title: Lower basal and postprandial muscle protein synthesis after 2 weeks single‐leg immobilization in older men: No protective effect of anti‐inflammatory medication
    Article Snippet: The primary antibodies from Cell Signaling Technology, Danvers, MA were total‐mTOR (no. 2983) diluted 1:1000, total‐p70‐S6K1 (no. 2708) diluted 1:1000, p‐p70‐S6K (Thr389; no. 9206) diluted 1:1000, total‐AKT1 (no. 2967) diluted 1:1000, p‐AKT (Thr308; no. 2965) diluted 1:1000, p‐eEF2 (Thr56; no. 2331) diluted 1:2000 and p‐4E‐BP1 (Thr37/46; no. 2855) diluted 1:1000, and from Abcam, Cambridge, MA total‐eEF2 (ab130187) diluted 1:500.

    Article Title: Rapamycin and mTORC2 inhibition synergistically reduce contraction‐stimulated muscle protein synthesis
    Article Snippet: Antibodies against total-mTOR (cat#2972), total-Rictor (cat#2114), total-Raptor (cat#2280), phospho-Akt (Thr308, cat#13038), phospho-Akt (Ser473, cat#9271), phospho-Akt1 (Ser473, cat#9018), phospho-Akt2 (Ser474, cat#8599), total-Akt (cat#9272), phospho-p70S6K (Thr389, cat#9205), total-p70S6K (cat#2708), phospho-rpS6 (Ser235/236, cat#2211), phospho-rpS6 (Ser240/244, cat#2215), total-rpS6 (cat#2217), phospho-PRAS40 (Ser183, cat#5936), phospho-PRAS40 (Thr246, cat#2640), total-PRAS40 (cat#2610), phospho-4E-BP1 (Thr37/46, cat#9459), phospho-4E-BP1 (Ser656, cat#9451), phospho-4E-BP1 (Thr70, cat#9455), total-4E-BP1 (cat#9452), phospho-AMPK (Thr172, cat#2535), total-AMPK (cat#2532), phospho-NDRG1 (Thr346, cat#5482), HK2 (cat#2106), and c-Myc (cat#5605) were obtained from Cell Signaling Technology This article is protected by copyright.

    Article Title: Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/mTOR Pathway
    Article Snippet: MIYAZAKI, M., M. NOGUCHI, and T. TAKEMASA.. Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/ mTOR Pathway.. Med.

    Article Title: Progranulin depletion inhibits proliferation via the transforming growth factor beta/SMAD family member 2 signaling axis in Kasumi-1 cells
    Article Snippet: Proteins were then transferred to PVDF membrane, which was blocked in 5% skim milk in TBST (Tris buffer saline with 0.1 % v/v Tween-20) at room temperature for 60 min. After washing with TBST buffer, the membrane was incubated with anti-progranulin (kind gift from Dr. Ginette Serrero; A&G Pharmaceutical, MD, USA), phospho-mTOR, total-mTOR, phospho-Akt, total Akt, phospho-ERK, total-ERK, cleaved PARP, caspase-3, Bax, Bcl-2, XIAP (Cell Signaling Technology, MA, USA), and β-actin (Sigma-Aldrich, MO, USA) antibodies at a concentration of 1.0 μg/mL (anti-progranulin), dilution of 1:15000 (anti-β-actin) or 1:1000 (other antibodies) in Can Get Signal (TOYOBO, Osaka, Japan) at 4 °C overnight.

    Article Title: A CCL2/ROS autoregulation loop is critical for cancer-associated fibroblasts-enhanced tumor growth of oral squamous cell carcinoma.
    Article Snippet: Antibodies were as follows: STAT3 (12640), c-Jun (9165), p-mTOR (5536), total-mTOR (2983), p-P70s6k (9234), total-P70s6k (2708), p-AKT (4060), total-AKT (4685) and Cyclin D (2978) were purchased from Cell Signaling Technology.

    Article Title: Homer Interactions Are Necessary for Metabotropic Glutamate Receptor-Induced Long-Term Depression and Translational Activation
    Article Snippet: Membranes were blocked in 5% nonfat dry milk and incubated with the following antibodies (Cell Signaling, Danvers, MA, unless otherwise noted) according to the manufacturer's instructions: total-ERK, phospho-ERK (Thr202/Tyr204), total-phosphoinositide-dependent kinase (PDK1), phospho-PDK1 (Ser 241), total-mTOR, phospho-mTOR (Ser2448), phospho-4EBP1 (Thr37/46), phospho-p70S6K (Thr389), elongation factor 1α (EF1α), and Actin (Millipore, Temecula, CA) at dilutions of 1:1000–1:5000.

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity
    Article Snippet: The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered saline-Tween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc-7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control.

    Multiple Displacement Amplification:

    Article Title: Resistance exercise enhances cognitive function in mouse.
    Article Snippet: ▼ Physical exercise has been shown to increase adult neurogenesis in the hippocampus and to enhance synaptic plasticity.. It has been demonstrated that these neuroprotective eff ects can be observed following aerobic exercise.. However, it remains unknown whether plasticity molecules, such as brain-derived neurotrophic factor (BDNF) and cyclic AMP response element-binding protein (CREB), are expressed in the hippocampus following resistance exercise.

    Article Title: Lower basal and postprandial muscle protein synthesis after 2 weeks single‐leg immobilization in older men: No protective effect of anti‐inflammatory medication
    Article Snippet: The primary antibodies from Cell Signaling Technology, Danvers, MA were total‐mTOR (no. 2983) diluted 1:1000, total‐p70‐S6K1 (no. 2708) diluted 1:1000, p‐p70‐S6K (Thr389; no. 9206) diluted 1:1000, total‐AKT1 (no. 2967) diluted 1:1000, p‐AKT (Thr308; no. 2965) diluted 1:1000, p‐eEF2 (Thr56; no. 2331) diluted 1:2000 and p‐4E‐BP1 (Thr37/46; no. 2855) diluted 1:1000, and from Abcam, Cambridge, MA total‐eEF2 (ab130187) diluted 1:500.

    Article Title: Rapamycin and mTORC2 inhibition synergistically reduce contraction‐stimulated muscle protein synthesis
    Article Snippet: Antibodies against total-mTOR (cat#2972), total-Rictor (cat#2114), total-Raptor (cat#2280), phospho-Akt (Thr308, cat#13038), phospho-Akt (Ser473, cat#9271), phospho-Akt1 (Ser473, cat#9018), phospho-Akt2 (Ser474, cat#8599), total-Akt (cat#9272), phospho-p70S6K (Thr389, cat#9205), total-p70S6K (cat#2708), phospho-rpS6 (Ser235/236, cat#2211), phospho-rpS6 (Ser240/244, cat#2215), total-rpS6 (cat#2217), phospho-PRAS40 (Ser183, cat#5936), phospho-PRAS40 (Thr246, cat#2640), total-PRAS40 (cat#2610), phospho-4E-BP1 (Thr37/46, cat#9459), phospho-4E-BP1 (Ser656, cat#9451), phospho-4E-BP1 (Thr70, cat#9455), total-4E-BP1 (cat#9452), phospho-AMPK (Thr172, cat#2535), total-AMPK (cat#2532), phospho-NDRG1 (Thr346, cat#5482), HK2 (cat#2106), and c-Myc (cat#5605) were obtained from Cell Signaling Technology This article is protected by copyright.

    Article Title: Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/mTOR Pathway
    Article Snippet: MIYAZAKI, M., M. NOGUCHI, and T. TAKEMASA.. Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/ mTOR Pathway.. Med.

    Article Title: Progranulin depletion inhibits proliferation via the transforming growth factor beta/SMAD family member 2 signaling axis in Kasumi-1 cells
    Article Snippet: Proteins were then transferred to PVDF membrane, which was blocked in 5% skim milk in TBST (Tris buffer saline with 0.1 % v/v Tween-20) at room temperature for 60 min. After washing with TBST buffer, the membrane was incubated with anti-progranulin (kind gift from Dr. Ginette Serrero; A&G Pharmaceutical, MD, USA), phospho-mTOR, total-mTOR, phospho-Akt, total Akt, phospho-ERK, total-ERK, cleaved PARP, caspase-3, Bax, Bcl-2, XIAP (Cell Signaling Technology, MA, USA), and β-actin (Sigma-Aldrich, MO, USA) antibodies at a concentration of 1.0 μg/mL (anti-progranulin), dilution of 1:15000 (anti-β-actin) or 1:1000 (other antibodies) in Can Get Signal (TOYOBO, Osaka, Japan) at 4 °C overnight.

    Article Title: A CCL2/ROS autoregulation loop is critical for cancer-associated fibroblasts-enhanced tumor growth of oral squamous cell carcinoma.
    Article Snippet: Antibodies were as follows: STAT3 (12640), c-Jun (9165), p-mTOR (5536), total-mTOR (2983), p-P70s6k (9234), total-P70s6k (2708), p-AKT (4060), total-AKT (4685) and Cyclin D (2978) were purchased from Cell Signaling Technology.

    Article Title: Homer Interactions Are Necessary for Metabotropic Glutamate Receptor-Induced Long-Term Depression and Translational Activation
    Article Snippet: Membranes were blocked in 5% nonfat dry milk and incubated with the following antibodies (Cell Signaling, Danvers, MA, unless otherwise noted) according to the manufacturer's instructions: total-ERK, phospho-ERK (Thr202/Tyr204), total-phosphoinositide-dependent kinase (PDK1), phospho-PDK1 (Ser 241), total-mTOR, phospho-mTOR (Ser2448), phospho-4EBP1 (Thr37/46), phospho-p70S6K (Thr389), elongation factor 1α (EF1α), and Actin (Millipore, Temecula, CA) at dilutions of 1:1000–1:5000.

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity
    Article Snippet: The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered saline-Tween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc-7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control.

    Control:

    Article Title: Resistance exercise enhances cognitive function in mouse.
    Article Snippet: ▼ Physical exercise has been shown to increase adult neurogenesis in the hippocampus and to enhance synaptic plasticity.. It has been demonstrated that these neuroprotective eff ects can be observed following aerobic exercise.. However, it remains unknown whether plasticity molecules, such as brain-derived neurotrophic factor (BDNF) and cyclic AMP response element-binding protein (CREB), are expressed in the hippocampus following resistance exercise.

    Article Title: Lower basal and postprandial muscle protein synthesis after 2 weeks single‐leg immobilization in older men: No protective effect of anti‐inflammatory medication
    Article Snippet: The primary antibodies from Cell Signaling Technology, Danvers, MA were total‐mTOR (no. 2983) diluted 1:1000, total‐p70‐S6K1 (no. 2708) diluted 1:1000, p‐p70‐S6K (Thr389; no. 9206) diluted 1:1000, total‐AKT1 (no. 2967) diluted 1:1000, p‐AKT (Thr308; no. 2965) diluted 1:1000, p‐eEF2 (Thr56; no. 2331) diluted 1:2000 and p‐4E‐BP1 (Thr37/46; no. 2855) diluted 1:1000, and from Abcam, Cambridge, MA total‐eEF2 (ab130187) diluted 1:500.

    Article Title: Rapamycin and mTORC2 inhibition synergistically reduce contraction‐stimulated muscle protein synthesis
    Article Snippet: Antibodies against total-mTOR (cat#2972), total-Rictor (cat#2114), total-Raptor (cat#2280), phospho-Akt (Thr308, cat#13038), phospho-Akt (Ser473, cat#9271), phospho-Akt1 (Ser473, cat#9018), phospho-Akt2 (Ser474, cat#8599), total-Akt (cat#9272), phospho-p70S6K (Thr389, cat#9205), total-p70S6K (cat#2708), phospho-rpS6 (Ser235/236, cat#2211), phospho-rpS6 (Ser240/244, cat#2215), total-rpS6 (cat#2217), phospho-PRAS40 (Ser183, cat#5936), phospho-PRAS40 (Thr246, cat#2640), total-PRAS40 (cat#2610), phospho-4E-BP1 (Thr37/46, cat#9459), phospho-4E-BP1 (Ser656, cat#9451), phospho-4E-BP1 (Thr70, cat#9455), total-4E-BP1 (cat#9452), phospho-AMPK (Thr172, cat#2535), total-AMPK (cat#2532), phospho-NDRG1 (Thr346, cat#5482), HK2 (cat#2106), and c-Myc (cat#5605) were obtained from Cell Signaling Technology This article is protected by copyright.

    Article Title: Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/mTOR Pathway
    Article Snippet: MIYAZAKI, M., M. NOGUCHI, and T. TAKEMASA.. Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/ mTOR Pathway.. Med.

    Article Title: Progranulin depletion inhibits proliferation via the transforming growth factor beta/SMAD family member 2 signaling axis in Kasumi-1 cells
    Article Snippet: Proteins were then transferred to PVDF membrane, which was blocked in 5% skim milk in TBST (Tris buffer saline with 0.1 % v/v Tween-20) at room temperature for 60 min. After washing with TBST buffer, the membrane was incubated with anti-progranulin (kind gift from Dr. Ginette Serrero; A&G Pharmaceutical, MD, USA), phospho-mTOR, total-mTOR, phospho-Akt, total Akt, phospho-ERK, total-ERK, cleaved PARP, caspase-3, Bax, Bcl-2, XIAP (Cell Signaling Technology, MA, USA), and β-actin (Sigma-Aldrich, MO, USA) antibodies at a concentration of 1.0 μg/mL (anti-progranulin), dilution of 1:15000 (anti-β-actin) or 1:1000 (other antibodies) in Can Get Signal (TOYOBO, Osaka, Japan) at 4 °C overnight.

    Article Title: A CCL2/ROS autoregulation loop is critical for cancer-associated fibroblasts-enhanced tumor growth of oral squamous cell carcinoma.
    Article Snippet: Antibodies were as follows: STAT3 (12640), c-Jun (9165), p-mTOR (5536), total-mTOR (2983), p-P70s6k (9234), total-P70s6k (2708), p-AKT (4060), total-AKT (4685) and Cyclin D (2978) were purchased from Cell Signaling Technology.

    Article Title: Homer Interactions Are Necessary for Metabotropic Glutamate Receptor-Induced Long-Term Depression and Translational Activation
    Article Snippet: Membranes were blocked in 5% nonfat dry milk and incubated with the following antibodies (Cell Signaling, Danvers, MA, unless otherwise noted) according to the manufacturer's instructions: total-ERK, phospho-ERK (Thr202/Tyr204), total-phosphoinositide-dependent kinase (PDK1), phospho-PDK1 (Ser 241), total-mTOR, phospho-mTOR (Ser2448), phospho-4EBP1 (Thr37/46), phospho-p70S6K (Thr389), elongation factor 1α (EF1α), and Actin (Millipore, Temecula, CA) at dilutions of 1:1000–1:5000.

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity
    Article Snippet: The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered saline-Tween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc-7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control.

    Membrane:

    Article Title: Resistance exercise enhances cognitive function in mouse.
    Article Snippet: ▼ Physical exercise has been shown to increase adult neurogenesis in the hippocampus and to enhance synaptic plasticity.. It has been demonstrated that these neuroprotective eff ects can be observed following aerobic exercise.. However, it remains unknown whether plasticity molecules, such as brain-derived neurotrophic factor (BDNF) and cyclic AMP response element-binding protein (CREB), are expressed in the hippocampus following resistance exercise.

    Article Title: Lower basal and postprandial muscle protein synthesis after 2 weeks single‐leg immobilization in older men: No protective effect of anti‐inflammatory medication
    Article Snippet: The primary antibodies from Cell Signaling Technology, Danvers, MA were total‐mTOR (no. 2983) diluted 1:1000, total‐p70‐S6K1 (no. 2708) diluted 1:1000, p‐p70‐S6K (Thr389; no. 9206) diluted 1:1000, total‐AKT1 (no. 2967) diluted 1:1000, p‐AKT (Thr308; no. 2965) diluted 1:1000, p‐eEF2 (Thr56; no. 2331) diluted 1:2000 and p‐4E‐BP1 (Thr37/46; no. 2855) diluted 1:1000, and from Abcam, Cambridge, MA total‐eEF2 (ab130187) diluted 1:500.

    Article Title: Rapamycin and mTORC2 inhibition synergistically reduce contraction‐stimulated muscle protein synthesis
    Article Snippet: Antibodies against total-mTOR (cat#2972), total-Rictor (cat#2114), total-Raptor (cat#2280), phospho-Akt (Thr308, cat#13038), phospho-Akt (Ser473, cat#9271), phospho-Akt1 (Ser473, cat#9018), phospho-Akt2 (Ser474, cat#8599), total-Akt (cat#9272), phospho-p70S6K (Thr389, cat#9205), total-p70S6K (cat#2708), phospho-rpS6 (Ser235/236, cat#2211), phospho-rpS6 (Ser240/244, cat#2215), total-rpS6 (cat#2217), phospho-PRAS40 (Ser183, cat#5936), phospho-PRAS40 (Thr246, cat#2640), total-PRAS40 (cat#2610), phospho-4E-BP1 (Thr37/46, cat#9459), phospho-4E-BP1 (Ser656, cat#9451), phospho-4E-BP1 (Thr70, cat#9455), total-4E-BP1 (cat#9452), phospho-AMPK (Thr172, cat#2535), total-AMPK (cat#2532), phospho-NDRG1 (Thr346, cat#5482), HK2 (cat#2106), and c-Myc (cat#5605) were obtained from Cell Signaling Technology This article is protected by copyright.

    Article Title: Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/mTOR Pathway
    Article Snippet: MIYAZAKI, M., M. NOGUCHI, and T. TAKEMASA.. Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/ mTOR Pathway.. Med.

    Article Title: Progranulin depletion inhibits proliferation via the transforming growth factor beta/SMAD family member 2 signaling axis in Kasumi-1 cells
    Article Snippet: Proteins were then transferred to PVDF membrane, which was blocked in 5% skim milk in TBST (Tris buffer saline with 0.1 % v/v Tween-20) at room temperature for 60 min. After washing with TBST buffer, the membrane was incubated with anti-progranulin (kind gift from Dr. Ginette Serrero; A&G Pharmaceutical, MD, USA), phospho-mTOR, total-mTOR, phospho-Akt, total Akt, phospho-ERK, total-ERK, cleaved PARP, caspase-3, Bax, Bcl-2, XIAP (Cell Signaling Technology, MA, USA), and β-actin (Sigma-Aldrich, MO, USA) antibodies at a concentration of 1.0 μg/mL (anti-progranulin), dilution of 1:15000 (anti-β-actin) or 1:1000 (other antibodies) in Can Get Signal (TOYOBO, Osaka, Japan) at 4 °C overnight.

    Article Title: A CCL2/ROS autoregulation loop is critical for cancer-associated fibroblasts-enhanced tumor growth of oral squamous cell carcinoma.
    Article Snippet: Antibodies were as follows: STAT3 (12640), c-Jun (9165), p-mTOR (5536), total-mTOR (2983), p-P70s6k (9234), total-P70s6k (2708), p-AKT (4060), total-AKT (4685) and Cyclin D (2978) were purchased from Cell Signaling Technology.

    Article Title: Homer Interactions Are Necessary for Metabotropic Glutamate Receptor-Induced Long-Term Depression and Translational Activation
    Article Snippet: Membranes were blocked in 5% nonfat dry milk and incubated with the following antibodies (Cell Signaling, Danvers, MA, unless otherwise noted) according to the manufacturer's instructions: total-ERK, phospho-ERK (Thr202/Tyr204), total-phosphoinositide-dependent kinase (PDK1), phospho-PDK1 (Ser 241), total-mTOR, phospho-mTOR (Ser2448), phospho-4EBP1 (Thr37/46), phospho-p70S6K (Thr389), elongation factor 1α (EF1α), and Actin (Millipore, Temecula, CA) at dilutions of 1:1000–1:5000.

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity
    Article Snippet: The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered saline-Tween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc-7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control.

    Concentration Assay:

    Article Title: Resistance exercise enhances cognitive function in mouse.
    Article Snippet: ▼ Physical exercise has been shown to increase adult neurogenesis in the hippocampus and to enhance synaptic plasticity.. It has been demonstrated that these neuroprotective eff ects can be observed following aerobic exercise.. However, it remains unknown whether plasticity molecules, such as brain-derived neurotrophic factor (BDNF) and cyclic AMP response element-binding protein (CREB), are expressed in the hippocampus following resistance exercise.

    Article Title: Lower basal and postprandial muscle protein synthesis after 2 weeks single‐leg immobilization in older men: No protective effect of anti‐inflammatory medication
    Article Snippet: The primary antibodies from Cell Signaling Technology, Danvers, MA were total‐mTOR (no. 2983) diluted 1:1000, total‐p70‐S6K1 (no. 2708) diluted 1:1000, p‐p70‐S6K (Thr389; no. 9206) diluted 1:1000, total‐AKT1 (no. 2967) diluted 1:1000, p‐AKT (Thr308; no. 2965) diluted 1:1000, p‐eEF2 (Thr56; no. 2331) diluted 1:2000 and p‐4E‐BP1 (Thr37/46; no. 2855) diluted 1:1000, and from Abcam, Cambridge, MA total‐eEF2 (ab130187) diluted 1:500.

    Article Title: Rapamycin and mTORC2 inhibition synergistically reduce contraction‐stimulated muscle protein synthesis
    Article Snippet: Antibodies against total-mTOR (cat#2972), total-Rictor (cat#2114), total-Raptor (cat#2280), phospho-Akt (Thr308, cat#13038), phospho-Akt (Ser473, cat#9271), phospho-Akt1 (Ser473, cat#9018), phospho-Akt2 (Ser474, cat#8599), total-Akt (cat#9272), phospho-p70S6K (Thr389, cat#9205), total-p70S6K (cat#2708), phospho-rpS6 (Ser235/236, cat#2211), phospho-rpS6 (Ser240/244, cat#2215), total-rpS6 (cat#2217), phospho-PRAS40 (Ser183, cat#5936), phospho-PRAS40 (Thr246, cat#2640), total-PRAS40 (cat#2610), phospho-4E-BP1 (Thr37/46, cat#9459), phospho-4E-BP1 (Ser656, cat#9451), phospho-4E-BP1 (Thr70, cat#9455), total-4E-BP1 (cat#9452), phospho-AMPK (Thr172, cat#2535), total-AMPK (cat#2532), phospho-NDRG1 (Thr346, cat#5482), HK2 (cat#2106), and c-Myc (cat#5605) were obtained from Cell Signaling Technology This article is protected by copyright.

    Article Title: Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/mTOR Pathway
    Article Snippet: MIYAZAKI, M., M. NOGUCHI, and T. TAKEMASA.. Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/ mTOR Pathway.. Med.

    Article Title: Progranulin depletion inhibits proliferation via the transforming growth factor beta/SMAD family member 2 signaling axis in Kasumi-1 cells
    Article Snippet: Proteins were then transferred to PVDF membrane, which was blocked in 5% skim milk in TBST (Tris buffer saline with 0.1 % v/v Tween-20) at room temperature for 60 min. After washing with TBST buffer, the membrane was incubated with anti-progranulin (kind gift from Dr. Ginette Serrero; A&G Pharmaceutical, MD, USA), phospho-mTOR, total-mTOR, phospho-Akt, total Akt, phospho-ERK, total-ERK, cleaved PARP, caspase-3, Bax, Bcl-2, XIAP (Cell Signaling Technology, MA, USA), and β-actin (Sigma-Aldrich, MO, USA) antibodies at a concentration of 1.0 μg/mL (anti-progranulin), dilution of 1:15000 (anti-β-actin) or 1:1000 (other antibodies) in Can Get Signal (TOYOBO, Osaka, Japan) at 4 °C overnight.

    Article Title: A CCL2/ROS autoregulation loop is critical for cancer-associated fibroblasts-enhanced tumor growth of oral squamous cell carcinoma.
    Article Snippet: Antibodies were as follows: STAT3 (12640), c-Jun (9165), p-mTOR (5536), total-mTOR (2983), p-P70s6k (9234), total-P70s6k (2708), p-AKT (4060), total-AKT (4685) and Cyclin D (2978) were purchased from Cell Signaling Technology.

    Article Title: Homer Interactions Are Necessary for Metabotropic Glutamate Receptor-Induced Long-Term Depression and Translational Activation
    Article Snippet: Membranes were blocked in 5% nonfat dry milk and incubated with the following antibodies (Cell Signaling, Danvers, MA, unless otherwise noted) according to the manufacturer's instructions: total-ERK, phospho-ERK (Thr202/Tyr204), total-phosphoinositide-dependent kinase (PDK1), phospho-PDK1 (Ser 241), total-mTOR, phospho-mTOR (Ser2448), phospho-4EBP1 (Thr37/46), phospho-p70S6K (Thr389), elongation factor 1α (EF1α), and Actin (Millipore, Temecula, CA) at dilutions of 1:1000–1:5000.

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity
    Article Snippet: The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered saline-Tween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc-7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control.

    Blocking Assay:

    Article Title: Resistance exercise enhances cognitive function in mouse.
    Article Snippet: ▼ Physical exercise has been shown to increase adult neurogenesis in the hippocampus and to enhance synaptic plasticity.. It has been demonstrated that these neuroprotective eff ects can be observed following aerobic exercise.. However, it remains unknown whether plasticity molecules, such as brain-derived neurotrophic factor (BDNF) and cyclic AMP response element-binding protein (CREB), are expressed in the hippocampus following resistance exercise.

    Article Title: Lower basal and postprandial muscle protein synthesis after 2 weeks single‐leg immobilization in older men: No protective effect of anti‐inflammatory medication
    Article Snippet: The primary antibodies from Cell Signaling Technology, Danvers, MA were total‐mTOR (no. 2983) diluted 1:1000, total‐p70‐S6K1 (no. 2708) diluted 1:1000, p‐p70‐S6K (Thr389; no. 9206) diluted 1:1000, total‐AKT1 (no. 2967) diluted 1:1000, p‐AKT (Thr308; no. 2965) diluted 1:1000, p‐eEF2 (Thr56; no. 2331) diluted 1:2000 and p‐4E‐BP1 (Thr37/46; no. 2855) diluted 1:1000, and from Abcam, Cambridge, MA total‐eEF2 (ab130187) diluted 1:500.

    Article Title: Rapamycin and mTORC2 inhibition synergistically reduce contraction‐stimulated muscle protein synthesis
    Article Snippet: Antibodies against total-mTOR (cat#2972), total-Rictor (cat#2114), total-Raptor (cat#2280), phospho-Akt (Thr308, cat#13038), phospho-Akt (Ser473, cat#9271), phospho-Akt1 (Ser473, cat#9018), phospho-Akt2 (Ser474, cat#8599), total-Akt (cat#9272), phospho-p70S6K (Thr389, cat#9205), total-p70S6K (cat#2708), phospho-rpS6 (Ser235/236, cat#2211), phospho-rpS6 (Ser240/244, cat#2215), total-rpS6 (cat#2217), phospho-PRAS40 (Ser183, cat#5936), phospho-PRAS40 (Thr246, cat#2640), total-PRAS40 (cat#2610), phospho-4E-BP1 (Thr37/46, cat#9459), phospho-4E-BP1 (Ser656, cat#9451), phospho-4E-BP1 (Thr70, cat#9455), total-4E-BP1 (cat#9452), phospho-AMPK (Thr172, cat#2535), total-AMPK (cat#2532), phospho-NDRG1 (Thr346, cat#5482), HK2 (cat#2106), and c-Myc (cat#5605) were obtained from Cell Signaling Technology This article is protected by copyright.

    Article Title: Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/mTOR Pathway
    Article Snippet: MIYAZAKI, M., M. NOGUCHI, and T. TAKEMASA.. Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/ mTOR Pathway.. Med.

    Article Title: Progranulin depletion inhibits proliferation via the transforming growth factor beta/SMAD family member 2 signaling axis in Kasumi-1 cells
    Article Snippet: Proteins were then transferred to PVDF membrane, which was blocked in 5% skim milk in TBST (Tris buffer saline with 0.1 % v/v Tween-20) at room temperature for 60 min. After washing with TBST buffer, the membrane was incubated with anti-progranulin (kind gift from Dr. Ginette Serrero; A&G Pharmaceutical, MD, USA), phospho-mTOR, total-mTOR, phospho-Akt, total Akt, phospho-ERK, total-ERK, cleaved PARP, caspase-3, Bax, Bcl-2, XIAP (Cell Signaling Technology, MA, USA), and β-actin (Sigma-Aldrich, MO, USA) antibodies at a concentration of 1.0 μg/mL (anti-progranulin), dilution of 1:15000 (anti-β-actin) or 1:1000 (other antibodies) in Can Get Signal (TOYOBO, Osaka, Japan) at 4 °C overnight.

    Article Title: A CCL2/ROS autoregulation loop is critical for cancer-associated fibroblasts-enhanced tumor growth of oral squamous cell carcinoma.
    Article Snippet: Antibodies were as follows: STAT3 (12640), c-Jun (9165), p-mTOR (5536), total-mTOR (2983), p-P70s6k (9234), total-P70s6k (2708), p-AKT (4060), total-AKT (4685) and Cyclin D (2978) were purchased from Cell Signaling Technology.

    Article Title: Homer Interactions Are Necessary for Metabotropic Glutamate Receptor-Induced Long-Term Depression and Translational Activation
    Article Snippet: Membranes were blocked in 5% nonfat dry milk and incubated with the following antibodies (Cell Signaling, Danvers, MA, unless otherwise noted) according to the manufacturer's instructions: total-ERK, phospho-ERK (Thr202/Tyr204), total-phosphoinositide-dependent kinase (PDK1), phospho-PDK1 (Ser 241), total-mTOR, phospho-mTOR (Ser2448), phospho-4EBP1 (Thr37/46), phospho-p70S6K (Thr389), elongation factor 1α (EF1α), and Actin (Millipore, Temecula, CA) at dilutions of 1:1000–1:5000.

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity
    Article Snippet: The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered saline-Tween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc-7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control.

    Activation Assay:

    Article Title: Resistance exercise enhances cognitive function in mouse.
    Article Snippet: ▼ Physical exercise has been shown to increase adult neurogenesis in the hippocampus and to enhance synaptic plasticity.. It has been demonstrated that these neuroprotective eff ects can be observed following aerobic exercise.. However, it remains unknown whether plasticity molecules, such as brain-derived neurotrophic factor (BDNF) and cyclic AMP response element-binding protein (CREB), are expressed in the hippocampus following resistance exercise.

    Article Title: Lower basal and postprandial muscle protein synthesis after 2 weeks single‐leg immobilization in older men: No protective effect of anti‐inflammatory medication
    Article Snippet: The primary antibodies from Cell Signaling Technology, Danvers, MA were total‐mTOR (no. 2983) diluted 1:1000, total‐p70‐S6K1 (no. 2708) diluted 1:1000, p‐p70‐S6K (Thr389; no. 9206) diluted 1:1000, total‐AKT1 (no. 2967) diluted 1:1000, p‐AKT (Thr308; no. 2965) diluted 1:1000, p‐eEF2 (Thr56; no. 2331) diluted 1:2000 and p‐4E‐BP1 (Thr37/46; no. 2855) diluted 1:1000, and from Abcam, Cambridge, MA total‐eEF2 (ab130187) diluted 1:500.

    Article Title: Rapamycin and mTORC2 inhibition synergistically reduce contraction‐stimulated muscle protein synthesis
    Article Snippet: Antibodies against total-mTOR (cat#2972), total-Rictor (cat#2114), total-Raptor (cat#2280), phospho-Akt (Thr308, cat#13038), phospho-Akt (Ser473, cat#9271), phospho-Akt1 (Ser473, cat#9018), phospho-Akt2 (Ser474, cat#8599), total-Akt (cat#9272), phospho-p70S6K (Thr389, cat#9205), total-p70S6K (cat#2708), phospho-rpS6 (Ser235/236, cat#2211), phospho-rpS6 (Ser240/244, cat#2215), total-rpS6 (cat#2217), phospho-PRAS40 (Ser183, cat#5936), phospho-PRAS40 (Thr246, cat#2640), total-PRAS40 (cat#2610), phospho-4E-BP1 (Thr37/46, cat#9459), phospho-4E-BP1 (Ser656, cat#9451), phospho-4E-BP1 (Thr70, cat#9455), total-4E-BP1 (cat#9452), phospho-AMPK (Thr172, cat#2535), total-AMPK (cat#2532), phospho-NDRG1 (Thr346, cat#5482), HK2 (cat#2106), and c-Myc (cat#5605) were obtained from Cell Signaling Technology This article is protected by copyright.

    Article Title: Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/mTOR Pathway
    Article Snippet: MIYAZAKI, M., M. NOGUCHI, and T. TAKEMASA.. Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/ mTOR Pathway.. Med.

    Article Title: Progranulin depletion inhibits proliferation via the transforming growth factor beta/SMAD family member 2 signaling axis in Kasumi-1 cells
    Article Snippet: Proteins were then transferred to PVDF membrane, which was blocked in 5% skim milk in TBST (Tris buffer saline with 0.1 % v/v Tween-20) at room temperature for 60 min. After washing with TBST buffer, the membrane was incubated with anti-progranulin (kind gift from Dr. Ginette Serrero; A&G Pharmaceutical, MD, USA), phospho-mTOR, total-mTOR, phospho-Akt, total Akt, phospho-ERK, total-ERK, cleaved PARP, caspase-3, Bax, Bcl-2, XIAP (Cell Signaling Technology, MA, USA), and β-actin (Sigma-Aldrich, MO, USA) antibodies at a concentration of 1.0 μg/mL (anti-progranulin), dilution of 1:15000 (anti-β-actin) or 1:1000 (other antibodies) in Can Get Signal (TOYOBO, Osaka, Japan) at 4 °C overnight.

    Article Title: A CCL2/ROS autoregulation loop is critical for cancer-associated fibroblasts-enhanced tumor growth of oral squamous cell carcinoma.
    Article Snippet: Antibodies were as follows: STAT3 (12640), c-Jun (9165), p-mTOR (5536), total-mTOR (2983), p-P70s6k (9234), total-P70s6k (2708), p-AKT (4060), total-AKT (4685) and Cyclin D (2978) were purchased from Cell Signaling Technology.

    Article Title: Homer Interactions Are Necessary for Metabotropic Glutamate Receptor-Induced Long-Term Depression and Translational Activation
    Article Snippet: Membranes were blocked in 5% nonfat dry milk and incubated with the following antibodies (Cell Signaling, Danvers, MA, unless otherwise noted) according to the manufacturer's instructions: total-ERK, phospho-ERK (Thr202/Tyr204), total-phosphoinositide-dependent kinase (PDK1), phospho-PDK1 (Ser 241), total-mTOR, phospho-mTOR (Ser2448), phospho-4EBP1 (Thr37/46), phospho-p70S6K (Thr389), elongation factor 1α (EF1α), and Actin (Millipore, Temecula, CA) at dilutions of 1:1000–1:5000.

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity
    Article Snippet: The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered saline-Tween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc-7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control.

    Scaffolding:

    Article Title: Resistance exercise enhances cognitive function in mouse.
    Article Snippet: ▼ Physical exercise has been shown to increase adult neurogenesis in the hippocampus and to enhance synaptic plasticity.. It has been demonstrated that these neuroprotective eff ects can be observed following aerobic exercise.. However, it remains unknown whether plasticity molecules, such as brain-derived neurotrophic factor (BDNF) and cyclic AMP response element-binding protein (CREB), are expressed in the hippocampus following resistance exercise.

    Article Title: Lower basal and postprandial muscle protein synthesis after 2 weeks single‐leg immobilization in older men: No protective effect of anti‐inflammatory medication
    Article Snippet: The primary antibodies from Cell Signaling Technology, Danvers, MA were total‐mTOR (no. 2983) diluted 1:1000, total‐p70‐S6K1 (no. 2708) diluted 1:1000, p‐p70‐S6K (Thr389; no. 9206) diluted 1:1000, total‐AKT1 (no. 2967) diluted 1:1000, p‐AKT (Thr308; no. 2965) diluted 1:1000, p‐eEF2 (Thr56; no. 2331) diluted 1:2000 and p‐4E‐BP1 (Thr37/46; no. 2855) diluted 1:1000, and from Abcam, Cambridge, MA total‐eEF2 (ab130187) diluted 1:500.

    Article Title: Rapamycin and mTORC2 inhibition synergistically reduce contraction‐stimulated muscle protein synthesis
    Article Snippet: Antibodies against total-mTOR (cat#2972), total-Rictor (cat#2114), total-Raptor (cat#2280), phospho-Akt (Thr308, cat#13038), phospho-Akt (Ser473, cat#9271), phospho-Akt1 (Ser473, cat#9018), phospho-Akt2 (Ser474, cat#8599), total-Akt (cat#9272), phospho-p70S6K (Thr389, cat#9205), total-p70S6K (cat#2708), phospho-rpS6 (Ser235/236, cat#2211), phospho-rpS6 (Ser240/244, cat#2215), total-rpS6 (cat#2217), phospho-PRAS40 (Ser183, cat#5936), phospho-PRAS40 (Thr246, cat#2640), total-PRAS40 (cat#2610), phospho-4E-BP1 (Thr37/46, cat#9459), phospho-4E-BP1 (Ser656, cat#9451), phospho-4E-BP1 (Thr70, cat#9455), total-4E-BP1 (cat#9452), phospho-AMPK (Thr172, cat#2535), total-AMPK (cat#2532), phospho-NDRG1 (Thr346, cat#5482), HK2 (cat#2106), and c-Myc (cat#5605) were obtained from Cell Signaling Technology This article is protected by copyright.

    Article Title: Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/mTOR Pathway
    Article Snippet: MIYAZAKI, M., M. NOGUCHI, and T. TAKEMASA.. Intermittent Reloading Attenuates Muscle Atrophy through Modulating Akt/ mTOR Pathway.. Med.

    Article Title: Progranulin depletion inhibits proliferation via the transforming growth factor beta/SMAD family member 2 signaling axis in Kasumi-1 cells
    Article Snippet: Proteins were then transferred to PVDF membrane, which was blocked in 5% skim milk in TBST (Tris buffer saline with 0.1 % v/v Tween-20) at room temperature for 60 min. After washing with TBST buffer, the membrane was incubated with anti-progranulin (kind gift from Dr. Ginette Serrero; A&G Pharmaceutical, MD, USA), phospho-mTOR, total-mTOR, phospho-Akt, total Akt, phospho-ERK, total-ERK, cleaved PARP, caspase-3, Bax, Bcl-2, XIAP (Cell Signaling Technology, MA, USA), and β-actin (Sigma-Aldrich, MO, USA) antibodies at a concentration of 1.0 μg/mL (anti-progranulin), dilution of 1:15000 (anti-β-actin) or 1:1000 (other antibodies) in Can Get Signal (TOYOBO, Osaka, Japan) at 4 °C overnight.

    Article Title: A CCL2/ROS autoregulation loop is critical for cancer-associated fibroblasts-enhanced tumor growth of oral squamous cell carcinoma.
    Article Snippet: Antibodies were as follows: STAT3 (12640), c-Jun (9165), p-mTOR (5536), total-mTOR (2983), p-P70s6k (9234), total-P70s6k (2708), p-AKT (4060), total-AKT (4685) and Cyclin D (2978) were purchased from Cell Signaling Technology.

    Article Title: Homer Interactions Are Necessary for Metabotropic Glutamate Receptor-Induced Long-Term Depression and Translational Activation
    Article Snippet: Membranes were blocked in 5% nonfat dry milk and incubated with the following antibodies (Cell Signaling, Danvers, MA, unless otherwise noted) according to the manufacturer's instructions: total-ERK, phospho-ERK (Thr202/Tyr204), total-phosphoinositide-dependent kinase (PDK1), phospho-PDK1 (Ser 241), total-mTOR, phospho-mTOR (Ser2448), phospho-4EBP1 (Thr37/46), phospho-p70S6K (Thr389), elongation factor 1α (EF1α), and Actin (Millipore, Temecula, CA) at dilutions of 1:1000–1:5000.

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity
    Article Snippet: The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered saline-Tween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc-7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control.



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    AMPAR-deficient CD4 T cells changed cytokine sensitivity and metabolically shifted toward glycolysis and lipid metabolism (A) Quantification of Foxp3 gMFI flow staining of WT and T ΔAMPAR iTregs cultured under the titration of IL-2 (mean ± SEM, ANOVA, representative of 3 independent experiments). (B) Representative flow plot of pSTAT5 + Foxp3 + expression (left) and quantification (right) from WT and T ΔAMPAR iTreg cultures (mean ± SEM, t test, representative of 3 independent experiments). (C) Quantification of CD4 + CD25 + frequency over the course of iTreg differentiation (mean ± SEM, ANOVA, representative of 3 independent experiments). (D) Western blot of WT and T ΔAMPAR CD4 T cells collected on day 1, 2, and 3 of iTreg differentiation, immunoblotted for phosphorylated <t>mTOR</t> (pmTOR), total mTOR, and β-Actin expression (top to bottom, representative of 3 independent experiments). (E) Quantification of CD71 + cells among CD4 + T cells on day 3 of iTreg differentiation. (mean ± SEM, t test, representative of 3 independent experiments). (F–I) Seahorse metabolic flux analysis of iTreg cells. (F) extracellular acidification rate (ECAR) measurement over time (minutes) and (G) quantification of glycolysis (left) and glycolytic capacity (right) measurements. (H) oxygen consumption rate (OCR) measurement over time (minutes) and (I) quantification of maximum respiratory capacity (left) and reserve capacity (right) measurements (mean ± SEM, ANOVA, representative of 3 independent experiments). (J and K) WT and T ΔAMPAR CD4 were cultured under iTreg polarizing conditions and subjected to bulk RNA sequencing and analysis. (J) GSEA plot of cholesterol metabolism pathway enrichment in T ΔAMPAR versus WT cells. (K) Heatmap of genes associated with cholesterol metabolism in T ΔAMPAR versus WT. (L) Quantification of Srebf1 , Srebf2 , Hmgcr , Acaca , Fasn , and Sqle mRNA relative expression in WT and T ΔAMPAR CD4 from iTreg culture determined by qPCR (mean ± SEM, ANOVA, representative of 2 independent experiments). (M) Western blot images (left) of WT and T ΔAMPAR immunoblotted for Srebf1 and Srebf2 (image is representative of 2 independent experiments) and quantification (right) of cleaved to precursor ratios analyzed by densitometry. (N and O) Cell staining for Lipid and cholesterol content. (N) Bodipy and (O) Filipin III gMFI flow staining of WT and T ΔAMPAR CD4 from day 2 iTreg cultures (mean ± SEM, t test, representative of 3 independent experiments). Statistical significance represented as ns > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.
    Antibodies Against Fibronectin, Claudin 1, Upa, Mt1 Mmp, Cleaved Caspase 3, Phospho Mtor, Total Mtor, And Phospho Akt, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    AMPAR-deficient CD4 T cells changed cytokine sensitivity and metabolically shifted toward glycolysis and lipid metabolism (A) Quantification of Foxp3 gMFI flow staining of WT and T ΔAMPAR iTregs cultured under the titration of IL-2 (mean ± SEM, ANOVA, representative of 3 independent experiments). (B) Representative flow plot of pSTAT5 + Foxp3 + expression (left) and quantification (right) from WT and T ΔAMPAR iTreg cultures (mean ± SEM, t test, representative of 3 independent experiments). (C) Quantification of CD4 + CD25 + frequency over the course of iTreg differentiation (mean ± SEM, ANOVA, representative of 3 independent experiments). (D) Western blot of WT and T ΔAMPAR CD4 T cells collected on day 1, 2, and 3 of iTreg differentiation, immunoblotted for phosphorylated <t>mTOR</t> (pmTOR), total mTOR, and β-Actin expression (top to bottom, representative of 3 independent experiments). (E) Quantification of CD71 + cells among CD4 + T cells on day 3 of iTreg differentiation. (mean ± SEM, t test, representative of 3 independent experiments). (F–I) Seahorse metabolic flux analysis of iTreg cells. (F) extracellular acidification rate (ECAR) measurement over time (minutes) and (G) quantification of glycolysis (left) and glycolytic capacity (right) measurements. (H) oxygen consumption rate (OCR) measurement over time (minutes) and (I) quantification of maximum respiratory capacity (left) and reserve capacity (right) measurements (mean ± SEM, ANOVA, representative of 3 independent experiments). (J and K) WT and T ΔAMPAR CD4 were cultured under iTreg polarizing conditions and subjected to bulk RNA sequencing and analysis. (J) GSEA plot of cholesterol metabolism pathway enrichment in T ΔAMPAR versus WT cells. (K) Heatmap of genes associated with cholesterol metabolism in T ΔAMPAR versus WT. (L) Quantification of Srebf1 , Srebf2 , Hmgcr , Acaca , Fasn , and Sqle mRNA relative expression in WT and T ΔAMPAR CD4 from iTreg culture determined by qPCR (mean ± SEM, ANOVA, representative of 2 independent experiments). (M) Western blot images (left) of WT and T ΔAMPAR immunoblotted for Srebf1 and Srebf2 (image is representative of 2 independent experiments) and quantification (right) of cleaved to precursor ratios analyzed by densitometry. (N and O) Cell staining for Lipid and cholesterol content. (N) Bodipy and (O) Filipin III gMFI flow staining of WT and T ΔAMPAR CD4 from day 2 iTreg cultures (mean ± SEM, t test, representative of 3 independent experiments). Statistical significance represented as ns > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.
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    AMPAR-deficient CD4 T cells changed cytokine sensitivity and metabolically shifted toward glycolysis and lipid metabolism (A) Quantification of Foxp3 gMFI flow staining of WT and T ΔAMPAR iTregs cultured under the titration of IL-2 (mean ± SEM, ANOVA, representative of 3 independent experiments). (B) Representative flow plot of pSTAT5 + Foxp3 + expression (left) and quantification (right) from WT and T ΔAMPAR iTreg cultures (mean ± SEM, t test, representative of 3 independent experiments). (C) Quantification of CD4 + CD25 + frequency over the course of iTreg differentiation (mean ± SEM, ANOVA, representative of 3 independent experiments). (D) Western blot of WT and T ΔAMPAR CD4 T cells collected on day 1, 2, and 3 of iTreg differentiation, immunoblotted for phosphorylated <t>mTOR</t> (pmTOR), total mTOR, and β-Actin expression (top to bottom, representative of 3 independent experiments). (E) Quantification of CD71 + cells among CD4 + T cells on day 3 of iTreg differentiation. (mean ± SEM, t test, representative of 3 independent experiments). (F–I) Seahorse metabolic flux analysis of iTreg cells. (F) extracellular acidification rate (ECAR) measurement over time (minutes) and (G) quantification of glycolysis (left) and glycolytic capacity (right) measurements. (H) oxygen consumption rate (OCR) measurement over time (minutes) and (I) quantification of maximum respiratory capacity (left) and reserve capacity (right) measurements (mean ± SEM, ANOVA, representative of 3 independent experiments). (J and K) WT and T ΔAMPAR CD4 were cultured under iTreg polarizing conditions and subjected to bulk RNA sequencing and analysis. (J) GSEA plot of cholesterol metabolism pathway enrichment in T ΔAMPAR versus WT cells. (K) Heatmap of genes associated with cholesterol metabolism in T ΔAMPAR versus WT. (L) Quantification of Srebf1 , Srebf2 , Hmgcr , Acaca , Fasn , and Sqle mRNA relative expression in WT and T ΔAMPAR CD4 from iTreg culture determined by qPCR (mean ± SEM, ANOVA, representative of 2 independent experiments). (M) Western blot images (left) of WT and T ΔAMPAR immunoblotted for Srebf1 and Srebf2 (image is representative of 2 independent experiments) and quantification (right) of cleaved to precursor ratios analyzed by densitometry. (N and O) Cell staining for Lipid and cholesterol content. (N) Bodipy and (O) Filipin III gMFI flow staining of WT and T ΔAMPAR CD4 from day 2 iTreg cultures (mean ± SEM, t test, representative of 3 independent experiments). Statistical significance represented as ns > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.
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    Image Search Results


    Current concepts of multiple amino acid–sensing input to mTORC1. AA, amino acid; Akt, RAC-α serine/threonine-protein kinase; ARF1, ADP-ribosylation factor 1; BCAA, branched-chain amino acid; CASTOR1, cytosolic arginine sensor for mTORC1 subunit 1; GATOR1/2, GAP activity toward RAGs 1/2; LARS, leucine-tRNA-synthase; LAT1/SLC7A5, L-type amino acid transporter 1; mTORC1, mechanistic target of rapamycin complex 1; PI3K, phosphatidylinositol-3-kinase; RAB1A, Ras-related protein Rab-1A; Rag A/B C/D, RAS-related GTP-binding protein A/B C/D; Ragulator, Late endosomal/lysosomal adaptor and MAPK and mTOR activator; Rheb, Ras homolog enriched in brain; SAM, S-adenosyl methionine; SAMTOR, S-adenosylmethionine sensor upstream of mTORC1; SAR1B, secretion-associated Ras-related GTPase 1B; Sestrin1/2, stress response protein 1/2; SLC38A9, solute carrier family 38 member 9; SNAT2, sodium-coupled neutral amino acid transporter 2; TARS2, threonyl-tRNA synthetase 2; TFEB, transcription factor EB; TSC1/2, tuberous sclerosis complex 1/2; V-ATPase, vacuolar H + -ATPase. Created in BioRender. Ramos dos Santos, A.C. (2026) https://BioRender.com/54b947z .

    Journal: The Journal of Nutrition

    Article Title: Amino Acid Signaling in Skeletal Muscle Is Blunted by Prematurity in a Piglet Model

    doi: 10.1016/j.tjnut.2025.101303

    Figure Lengend Snippet: Current concepts of multiple amino acid–sensing input to mTORC1. AA, amino acid; Akt, RAC-α serine/threonine-protein kinase; ARF1, ADP-ribosylation factor 1; BCAA, branched-chain amino acid; CASTOR1, cytosolic arginine sensor for mTORC1 subunit 1; GATOR1/2, GAP activity toward RAGs 1/2; LARS, leucine-tRNA-synthase; LAT1/SLC7A5, L-type amino acid transporter 1; mTORC1, mechanistic target of rapamycin complex 1; PI3K, phosphatidylinositol-3-kinase; RAB1A, Ras-related protein Rab-1A; Rag A/B C/D, RAS-related GTP-binding protein A/B C/D; Ragulator, Late endosomal/lysosomal adaptor and MAPK and mTOR activator; Rheb, Ras homolog enriched in brain; SAM, S-adenosyl methionine; SAMTOR, S-adenosylmethionine sensor upstream of mTORC1; SAR1B, secretion-associated Ras-related GTPase 1B; Sestrin1/2, stress response protein 1/2; SLC38A9, solute carrier family 38 member 9; SNAT2, sodium-coupled neutral amino acid transporter 2; TARS2, threonyl-tRNA synthetase 2; TFEB, transcription factor EB; TSC1/2, tuberous sclerosis complex 1/2; V-ATPase, vacuolar H + -ATPase. Created in BioRender. Ramos dos Santos, A.C. (2026) https://BioRender.com/54b947z .

    Article Snippet: Immunoblotting and immunoprecipitation assays were performed using the following primary antibodies: ARF1 (Cat. No. 68069-1-AP; ProteinTech Group), CASTOR1/GATSL3 (Cat. No. A13309; Boster Biological Technology), LARS (Cat. No. A304-316A; Bethyl Laboratories), Mios, a GATOR2 subunit (Cat. No. 13557; Cell Signaling Technology), mTOR phosphorylated Ser2448 (Cat. No. 2971; Cell Signaling Technology), mTOR total (Cat. No. 2972; Cell Signaling Technology), NPRL2, a GATOR1 subunit (Cat. No. 37344, 37344; Cell Signaling Technology), Rab1A (Cat. No. 13075; Cell Signaling Technology), RagA (Cat. No. 4357; Cell Signaling Technology), RagC (Cat. No. 9480; Cell Signaling Technology), Raptor (Cat. No. 2280; Cell Signaling Technology), SAMTOR/BMT2 (Cat. No. 21744-1-AP; ProteinTech Group), SAR1B (Cat. No. 22292-1-AP; ProteinTech Group), Sestrin1 (Cat. No. 55010-1-AP; ProteinTech Group), Sestrin2 (Cat. No. 66297-1-Ig; ProteinTech Group), SLC38A2/sodium-coupled neutral amino acid transporter (SNAT)2 (Cat. No. ARP33059_P050; Aviva Systems Biology), SLC38A9 (Cat. No. AAS59532C; Antibody Verify), LAT/1SLC7A5 (Cat. No. BMP011; MBL International), and TARS2 (Cat. No. 15067-1-AP; ProteinTech Group).

    Techniques: Activity Assay, Binding Assay

    AMPAR-deficient CD4 T cells changed cytokine sensitivity and metabolically shifted toward glycolysis and lipid metabolism (A) Quantification of Foxp3 gMFI flow staining of WT and T ΔAMPAR iTregs cultured under the titration of IL-2 (mean ± SEM, ANOVA, representative of 3 independent experiments). (B) Representative flow plot of pSTAT5 + Foxp3 + expression (left) and quantification (right) from WT and T ΔAMPAR iTreg cultures (mean ± SEM, t test, representative of 3 independent experiments). (C) Quantification of CD4 + CD25 + frequency over the course of iTreg differentiation (mean ± SEM, ANOVA, representative of 3 independent experiments). (D) Western blot of WT and T ΔAMPAR CD4 T cells collected on day 1, 2, and 3 of iTreg differentiation, immunoblotted for phosphorylated mTOR (pmTOR), total mTOR, and β-Actin expression (top to bottom, representative of 3 independent experiments). (E) Quantification of CD71 + cells among CD4 + T cells on day 3 of iTreg differentiation. (mean ± SEM, t test, representative of 3 independent experiments). (F–I) Seahorse metabolic flux analysis of iTreg cells. (F) extracellular acidification rate (ECAR) measurement over time (minutes) and (G) quantification of glycolysis (left) and glycolytic capacity (right) measurements. (H) oxygen consumption rate (OCR) measurement over time (minutes) and (I) quantification of maximum respiratory capacity (left) and reserve capacity (right) measurements (mean ± SEM, ANOVA, representative of 3 independent experiments). (J and K) WT and T ΔAMPAR CD4 were cultured under iTreg polarizing conditions and subjected to bulk RNA sequencing and analysis. (J) GSEA plot of cholesterol metabolism pathway enrichment in T ΔAMPAR versus WT cells. (K) Heatmap of genes associated with cholesterol metabolism in T ΔAMPAR versus WT. (L) Quantification of Srebf1 , Srebf2 , Hmgcr , Acaca , Fasn , and Sqle mRNA relative expression in WT and T ΔAMPAR CD4 from iTreg culture determined by qPCR (mean ± SEM, ANOVA, representative of 2 independent experiments). (M) Western blot images (left) of WT and T ΔAMPAR immunoblotted for Srebf1 and Srebf2 (image is representative of 2 independent experiments) and quantification (right) of cleaved to precursor ratios analyzed by densitometry. (N and O) Cell staining for Lipid and cholesterol content. (N) Bodipy and (O) Filipin III gMFI flow staining of WT and T ΔAMPAR CD4 from day 2 iTreg cultures (mean ± SEM, t test, representative of 3 independent experiments). Statistical significance represented as ns > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

    Journal: iScience

    Article Title: The ionotropic AMPA receptor contributes to autoimmunity via altered regulatory T cell differentiation

    doi: 10.1016/j.isci.2025.114267

    Figure Lengend Snippet: AMPAR-deficient CD4 T cells changed cytokine sensitivity and metabolically shifted toward glycolysis and lipid metabolism (A) Quantification of Foxp3 gMFI flow staining of WT and T ΔAMPAR iTregs cultured under the titration of IL-2 (mean ± SEM, ANOVA, representative of 3 independent experiments). (B) Representative flow plot of pSTAT5 + Foxp3 + expression (left) and quantification (right) from WT and T ΔAMPAR iTreg cultures (mean ± SEM, t test, representative of 3 independent experiments). (C) Quantification of CD4 + CD25 + frequency over the course of iTreg differentiation (mean ± SEM, ANOVA, representative of 3 independent experiments). (D) Western blot of WT and T ΔAMPAR CD4 T cells collected on day 1, 2, and 3 of iTreg differentiation, immunoblotted for phosphorylated mTOR (pmTOR), total mTOR, and β-Actin expression (top to bottom, representative of 3 independent experiments). (E) Quantification of CD71 + cells among CD4 + T cells on day 3 of iTreg differentiation. (mean ± SEM, t test, representative of 3 independent experiments). (F–I) Seahorse metabolic flux analysis of iTreg cells. (F) extracellular acidification rate (ECAR) measurement over time (minutes) and (G) quantification of glycolysis (left) and glycolytic capacity (right) measurements. (H) oxygen consumption rate (OCR) measurement over time (minutes) and (I) quantification of maximum respiratory capacity (left) and reserve capacity (right) measurements (mean ± SEM, ANOVA, representative of 3 independent experiments). (J and K) WT and T ΔAMPAR CD4 were cultured under iTreg polarizing conditions and subjected to bulk RNA sequencing and analysis. (J) GSEA plot of cholesterol metabolism pathway enrichment in T ΔAMPAR versus WT cells. (K) Heatmap of genes associated with cholesterol metabolism in T ΔAMPAR versus WT. (L) Quantification of Srebf1 , Srebf2 , Hmgcr , Acaca , Fasn , and Sqle mRNA relative expression in WT and T ΔAMPAR CD4 from iTreg culture determined by qPCR (mean ± SEM, ANOVA, representative of 2 independent experiments). (M) Western blot images (left) of WT and T ΔAMPAR immunoblotted for Srebf1 and Srebf2 (image is representative of 2 independent experiments) and quantification (right) of cleaved to precursor ratios analyzed by densitometry. (N and O) Cell staining for Lipid and cholesterol content. (N) Bodipy and (O) Filipin III gMFI flow staining of WT and T ΔAMPAR CD4 from day 2 iTreg cultures (mean ± SEM, t test, representative of 3 independent experiments). Statistical significance represented as ns > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

    Article Snippet: Probes included: anti-pSMAD3 (Cell signaling, 9520), anti-total SMAD2/3 (Cell signaling, 3102), anti-pmTOR (Cell signaling, 2971), anti-total mTOR (Cell signaling, 2983), anti-Srebp1 (Abcam, ab3259), anti-Srebp2 (Abcam, ab30682), anti-β-tubulin (Cell signaling, 86298) and anti-β-actin (Cell signaling, 4970).

    Techniques: Metabolic Labelling, Staining, Cell Culture, Titration, Expressing, Western Blot, RNA Sequencing