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akt substrate 160  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc akt substrate 160
    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 <t>AS160</t> 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 <t>S318,</t> 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.
    Akt Substrate 160, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/akt+substrate+160/Phospho-AS160+(Ser318)+Rabbit+mAb/pmc12816905-99-63-69
    Average 93 stars, based on 31 article reviews
    akt substrate 160 - by Bioz Stars, 2026-09
    93/100 stars

    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

    Western Blot:

    Article Title: Growth hormone (GH)-induced insulin resistance is rapidly reversible: an experimental study in GH-deficient adults.
    Article Snippet: .. Immunoblotting was performed using primary antibodies as follows: phospho-signal transducer and activator of transcription 5 (Stat5), Stat5, phosphoAktser473, Akt2, phospho-Akt substrate (PAS), and Akt substrate 160 (AS160), all obtained from Cell Signaling (Beverly, MA). .. Membranes were incubated with horseradish peroxidase-coupled secondary antibodies (GE Healthcare, Waukesha, WI), visualized by SuperSignal West Dura chemiluminescent substrate (Thermo Fisher Scientific, Waltham, MA), and quantified by the UVP BioImaging System (Upland, CA).

    Article Title: Weightlifting outperforms voluntary wheel running for improving adiposity and insulin sensitivity in obese mice.
    Article Snippet: .. Standard insulin treatment and immunoblotting procedures were performed as previously described.26,30 The plantaris was chosen based on previous studies showing efficacious response to exercise training as well as insulin response.30 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). .. Secondary antibodies were goat anti-rabbit IR800 and anti-mouse IR680 (# 926-32211 and 926-32222 respectively; LiCor Biosciences; Lincoln, NE, USA).

    Article Title: Insulin and GH Signaling in Human Skeletal Muscle In Vivo following Exogenous GH Exposure: Impact of an Oral Glucose Load
    Article Snippet: .. Immunoblotting was performed using primary antibodies as follows: phosho-STAT5, STAT5, phosho Akt, Akt2, phospho-P38, P38, phosho-Akt substrate (PAS), and Akt substrate 160 (AS160), all obtained from Cell Signaling (Beverly, MA). .. Membranes were incubated with horseradish peroxidase–coupled secondary antibodies, visualized by BioWest enhanced chemiluminescence (UVP LabWorks, Upland, CA) and quantified by the UVP BioImaging System.

    Membrane:

    Article Title: Alteration of cardiac glucose metabolism in association to low birth weight: experimental evidence in lambs with left ventricular hypertrophy.
    Article Snippet: .. Equal amounts of extracted protein were subjected to SDS-PAGE, transferred to a membrane as previously described [29,30] and then incubated with primary antibody: carnitine palmitoyltransferase-I (CPT-I; Santa Cruz Biotechnology, CA, USA), acetyl CoA carboxylase (ACC Cell Signalling Technology, MA, USA), phospho-ACC (Ser79; Cell Signalling Technology), insulin receptor (IR; Abcam,Cambridge, UK), phospho-IR (Tyr1162/1163; Santa Cruz Biotechnology), phospho-protein kinase B (Akt, Ser473; Cell Signalling Technology), Akt-1 (Cell Signalling Technology), Akt-2 (Cell Signalling Technology), Akt substrate 160 (AS160; Cell Signalling Technology), phospho-AS160 (Thr642; Cell Signalling Technology), glucose transporter (GLUT)-4 (Abcam), GLUT-1 (Santa Cruz Biotechnology), glycogen synthase kinase (GSK)-3α/β (Santa Cruz Biotechnology), phospho-GSK-3β (Ser9; Santa Cruz Biotechnology), glycogen synthase (GS; Cell Signalling Technology), phospho-GS (Ser641; Cell Signalling Technology) and pyruvate dehydrogenase kinase-4 (PDK-4; Abcam) overnight at 4 °C with agitation and detected by enhanced chemiluminescence as previously described [30]. ..

    Incubation:

    Article Title: Alteration of cardiac glucose metabolism in association to low birth weight: experimental evidence in lambs with left ventricular hypertrophy.
    Article Snippet: .. Equal amounts of extracted protein were subjected to SDS-PAGE, transferred to a membrane as previously described [29,30] and then incubated with primary antibody: carnitine palmitoyltransferase-I (CPT-I; Santa Cruz Biotechnology, CA, USA), acetyl CoA carboxylase (ACC Cell Signalling Technology, MA, USA), phospho-ACC (Ser79; Cell Signalling Technology), insulin receptor (IR; Abcam,Cambridge, UK), phospho-IR (Tyr1162/1163; Santa Cruz Biotechnology), phospho-protein kinase B (Akt, Ser473; Cell Signalling Technology), Akt-1 (Cell Signalling Technology), Akt-2 (Cell Signalling Technology), Akt substrate 160 (AS160; Cell Signalling Technology), phospho-AS160 (Thr642; Cell Signalling Technology), glucose transporter (GLUT)-4 (Abcam), GLUT-1 (Santa Cruz Biotechnology), glycogen synthase kinase (GSK)-3α/β (Santa Cruz Biotechnology), phospho-GSK-3β (Ser9; Santa Cruz Biotechnology), glycogen synthase (GS; Cell Signalling Technology), phospho-GS (Ser641; Cell Signalling Technology) and pyruvate dehydrogenase kinase-4 (PDK-4; Abcam) overnight at 4 °C with agitation and detected by enhanced chemiluminescence as previously described [30]. ..

    Ubiquitin Proteomics:

    Article Title: Weightlifting outperforms voluntary wheel running for improving adiposity and insulin sensitivity in obese mice
    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). .. Secondary antibodies were goat anti-rabbit IR800 and anti-mouse IR680 (# 926-32211 and 926-32222 respectively; LiCor Biosciences; Lincoln, NE, USA).

    Article Title: Weightlifting outperforms voluntary wheel running for improving adiposity and insulin sensitivity in obese mice.
    Article Snippet: .. Standard insulin treatment and immunoblotting procedures were performed as previously described.26,30 The plantaris was chosen based on previous studies showing efficacious response to exercise training as well as insulin response.30 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). .. Secondary antibodies were goat anti-rabbit IR800 and anti-mouse IR680 (# 926-32211 and 926-32222 respectively; LiCor Biosciences; Lincoln, NE, USA).

    Binding Assay:

    Article Title: Weightlifting outperforms voluntary wheel running for improving adiposity and insulin sensitivity in obese mice
    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). .. Secondary antibodies were goat anti-rabbit IR800 and anti-mouse IR680 (# 926-32211 and 926-32222 respectively; LiCor Biosciences; Lincoln, NE, USA).

    Article Title: Weightlifting outperforms voluntary wheel running for improving adiposity and insulin sensitivity in obese mice.
    Article Snippet: .. Standard insulin treatment and immunoblotting procedures were performed as previously described.26,30 The plantaris was chosen based on previous studies showing efficacious response to exercise training as well as insulin response.30 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). .. Secondary antibodies were goat anti-rabbit IR800 and anti-mouse IR680 (# 926-32211 and 926-32222 respectively; LiCor Biosciences; Lincoln, NE, USA).



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    Figure 2. T4 increased <t>AS160</t> phosphorylation in heart. Mean ± SD values for (A) insulin receptor protein expression (IR), (B) Insulin receptor substrate -1 (IRS) protein expression , (C) Phosphoinositide 3-kinases p110 (PI3Kp110) protein expression, (D) protein kinase B (AKT) protein expression, (E) phosphorylated AKT (p-AKT) protein expression, (F) p-AKT/AKT ratio, (G) phosphorylated AMP-activated protein kinase (p-AMPK) protein expression, (H) <t>AKT</t> <t>Substrate</t> <t>160</t> (AS160) protein expression, (I) p-AS160 protein expression, (J) p-AS160/AS160 ratio, and (K) representative blot of LETO, LETO + T4, OLETF, and OLETF + T4 rats (n=6-7) PS, ponceau stain. *, Significant difference from LETO p<0.05. &, Significant difference from OLETF p<0.05. $, Significant difference from LETO + T4 p<0.05. #, Significant difference from OLETF + T4 p<0.05.
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    anti-akt substrate of 160 kda (as160; #abs54)  (Millipore)
    90
    Millipore anti-akt substrate of 160 kda (as160; #abs54)
    Figure 2. T4 increased <t>AS160</t> phosphorylation in heart. Mean ± SD values for (A) insulin receptor protein expression (IR), (B) Insulin receptor substrate -1 (IRS) protein expression , (C) Phosphoinositide 3-kinases p110 (PI3Kp110) protein expression, (D) protein kinase B (AKT) protein expression, (E) phosphorylated AKT (p-AKT) protein expression, (F) p-AKT/AKT ratio, (G) phosphorylated AMP-activated protein kinase (p-AMPK) protein expression, (H) <t>AKT</t> <t>Substrate</t> <t>160</t> (AS160) protein expression, (I) p-AS160 protein expression, (J) p-AS160/AS160 ratio, and (K) representative blot of LETO, LETO + T4, OLETF, and OLETF + T4 rats (n=6-7) PS, ponceau stain. *, Significant difference from LETO p<0.05. &, Significant difference from OLETF p<0.05. $, Significant difference from LETO + T4 p<0.05. #, Significant difference from OLETF + T4 p<0.05.
    Anti Akt Substrate Of 160 Kda (As160; #Abs54), supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/akt+substrate+160/as160+antibody/pmc09648397-43-0-31
    Average 90 stars, based on 1 article reviews
    anti-akt substrate of 160 kda (as160; #abs54) - by Bioz Stars, 2026-09
    90/100 stars
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    anti-akt substrate of 160 kda as160  (Millipore)
    90
    Millipore anti-akt substrate of 160 kda as160
    Figure 2. T4 increased <t>AS160</t> phosphorylation in heart. Mean ± SD values for (A) insulin receptor protein expression (IR), (B) Insulin receptor substrate -1 (IRS) protein expression , (C) Phosphoinositide 3-kinases p110 (PI3Kp110) protein expression, (D) protein kinase B (AKT) protein expression, (E) phosphorylated AKT (p-AKT) protein expression, (F) p-AKT/AKT ratio, (G) phosphorylated AMP-activated protein kinase (p-AMPK) protein expression, (H) <t>AKT</t> <t>Substrate</t> <t>160</t> (AS160) protein expression, (I) p-AS160 protein expression, (J) p-AS160/AS160 ratio, and (K) representative blot of LETO, LETO + T4, OLETF, and OLETF + T4 rats (n=6-7) PS, ponceau stain. *, Significant difference from LETO p<0.05. &, Significant difference from OLETF p<0.05. $, Significant difference from LETO + T4 p<0.05. #, Significant difference from OLETF + T4 p<0.05.
    Anti Akt Substrate Of 160 Kda As160, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/akt+substrate+160/as160+antibody/pmc09359348-32-0-9
    Average 90 stars, based on 1 article reviews
    anti-akt substrate of 160 kda as160 - by Bioz Stars, 2026-09
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    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

    Figure 2. T4 increased AS160 phosphorylation in heart. Mean ± SD values for (A) insulin receptor protein expression (IR), (B) Insulin receptor substrate -1 (IRS) protein expression , (C) Phosphoinositide 3-kinases p110 (PI3Kp110) protein expression, (D) protein kinase B (AKT) protein expression, (E) phosphorylated AKT (p-AKT) protein expression, (F) p-AKT/AKT ratio, (G) phosphorylated AMP-activated protein kinase (p-AMPK) protein expression, (H) AKT Substrate 160 (AS160) protein expression, (I) p-AS160 protein expression, (J) p-AS160/AS160 ratio, and (K) representative blot of LETO, LETO + T4, OLETF, and OLETF + T4 rats (n=6-7) PS, ponceau stain. *, Significant difference from LETO p<0.05. &, Significant difference from OLETF p<0.05. $, Significant difference from LETO + T4 p<0.05. #, Significant difference from OLETF + T4 p<0.05.

    Journal: Molecular and cellular endocrinology

    Article Title: Exogenous Thyroxine Increases Cardiac GLUT4 Translocation in Insulin Resistant OLETF Rats.

    doi: 10.1016/j.mce.2024.112254

    Figure Lengend Snippet: Figure 2. T4 increased AS160 phosphorylation in heart. Mean ± SD values for (A) insulin receptor protein expression (IR), (B) Insulin receptor substrate -1 (IRS) protein expression , (C) Phosphoinositide 3-kinases p110 (PI3Kp110) protein expression, (D) protein kinase B (AKT) protein expression, (E) phosphorylated AKT (p-AKT) protein expression, (F) p-AKT/AKT ratio, (G) phosphorylated AMP-activated protein kinase (p-AMPK) protein expression, (H) AKT Substrate 160 (AS160) protein expression, (I) p-AS160 protein expression, (J) p-AS160/AS160 ratio, and (K) representative blot of LETO, LETO + T4, OLETF, and OLETF + T4 rats (n=6-7) PS, ponceau stain. *, Significant difference from LETO p<0.05. &, Significant difference from OLETF p<0.05. $, Significant difference from LETO + T4 p<0.05. #, Significant difference from OLETF + T4 p<0.05.

    Article Snippet: 126 Membranes were incubated with primary antibodies against GLUT4 (Abcam, ab33780, Boston, 127 MA, USA), pS488-GLUT4 (Abcam, ab188317, Boston, MA, USA), insulin receptor (IR) (Cell 128 Signaling Technology, 3025S, USA), insulin receptor substrate (IRS) 1 + IRS 2 (abcam, ab40777, 129 Boston, MA, USA), phosphoinositide 3-kinases p110 (PI3K)p110 (Proteintech, 67071-1-Ig, 130 Rosemont, IL, USA), protein kinase B (AKT) (Cell Signaling Technology, 9272S, USA), p-AKT (Cell 131 Jo urn al Pr e-p roo f REVISED Manuscript (text UNmarked) 7 Signaling Technology, 4060S, USA), anti-AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho 132 T172) (Abcam, ab133448, Boston, MA, USA), AKT substrate 160 (AS160) (C69A7) (Cell Signaling 133 Technology, 2670S, USA), Phospho-AS160 (Thr642) (D27E6) (Cell Signaling Technology, 8881S, 134 USA), hexokinase 2 (Proteintech, 22029-1-AP, Rosemont, IL, USA), phosphofructokinase muscle 135 type (PFKM) (Abcam, ab154804, Boston, MA, USA), monocarboxylate transporter 8 (MCT8) 136 (Abcam, ab302706, Boston, MA, USA), Type II iodothyronine deiodinase (DIO2) (Proteintech, 137 26513-1-AP, Rosemont, IL, USA), thyroid hormone receptor β (TRβ1) (Abcam, ab180612, Boston, 138 MA, USA), thyroid hormone receptor alpha (THRa) (Proteintech, 66703-1-Ig, Rosemont, IL, USA), 139 the fatty acid transporter, cluster of differentiation 36 (CD36) (Thermo Fisher, PA1-16813, 140 Houston, TX, USA), fatty acid transporter (FATP1) (MyBioSource, MBS9384813, San Diego, CA, 141 USA), carnitine palmitoyltransferase 2 (CPT2) (Invitrogen, PA5-30420, Waltham, MA, USA), acyl-142 coA oxidase 1 (ACOX) (Thermo Fisher, PA5-76341, USA), acetyl-coA carboxylase 1 (ACC) (Cell 143 Signaling Technology, 3662S, USA), diacylglycerol O-acyltransferase 1 (DGAT1) (Thermo Fisher, 144 PA5-79150, Houston, TX, USA), and glycerol-3-phosphate acyltransferase (GPAM) (abcam, 145 ab69990, Boston, MA, USA).

    Techniques: Phospho-proteomics, Expressing, Staining