akt substrate 160  (Cell Signaling Technology Inc)

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

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