s473  (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: Journal of Sport and Health Science

Article Title: Exercise training-induced extracellular miR-136-3p modulates glucose uptake and myogenesis through targeting of NRDC in human skeletal muscle

doi: 10.1016/j.jshs.2025.101091

Figure Lengend Snippet: miR-136-3p-induced increase in glucose uptake is independent of changes in canonical GLUT4 signaling pathways. The mRNA expression of GLUT4, TBC1D4, and AMPKα in primary human myotubes (A) transfected with miR-136-3p. Representative immunoblot and quantification of (B) AMPKα, (C) TBC1D4, (D) P-TBC1D4, (E) AKT, and (F) P-AKT in primary human myotubes transfected with miR-136-3p and subsequently incubated under basal or insulin-stimulated (120 nM, 1 h) conditions. Insets show representative Western blot image and total ponceau staining for loading control. Results are expressed as mean ± standard error of the mean. * p < 0.05, ** p < 0.005. AKT = protein kinase B; AMPKα = AMP-activated protein kinase α; GLUT4 = glucose transporter 4; miR = microRNA; NC = negative control; ns = no significance; P-AKT = phosphorylated AKT; P-TBC1D4 = phosphorylated TBC1D4; TBC1D4 = Tre-2/BUB2/CDC16 domain family member 4.

Article Snippet: Membranes were incubated with primary antibodies directed to NRDC (sc-137199; Santa Cruz Biotechnology, Dallas, TX, USA), AMP-activated protein kinase α (AMPKα) (#2532; Cell Signaling Technology, Danvers, MA, USA), Akt substrate of 160 kDa (AS160) (#2670; Cell Signaling Technology), phospho-AS160 (#8730; Cell Signaling Technology), protein kinase B (AKT) (#9272; Cell Signaling Technology), phospho-AKT (Ser473) (#9271; Cell Signaling Technology), or puromycin (MABE343; Merck Burlington, MA, USA).

Techniques: Protein-Protein interactions, Expressing, Transfection, Western Blot, Incubation, Staining, Control, Negative Control

Journal: Journal of Animal Science and Biotechnology

Article Title: ZBED6 deletion activates FNDC1 -mediated PI3K/AKT and MAPK/ERK1/2 signaling pathways to promote myogenesis in pigs

doi: 10.1186/s40104-026-01504-w

Figure Lengend Snippet: Antibodies used in this study

Article Snippet: p-AKT Ser473 , Proteintech , 66444-1-Ig.

Techniques: Cell-Signaling

Journal: Chemistry & Biodiversity

Article Title: Network Pharmacology and In Vivo Evaluation of Modified Huanglian Wendan Decoction in Polycystic Ovary Syndrome With Insulin Resistance: Association With AGE–RAGE‐Related Signaling

doi: 10.1002/cbdv.71737

Figure Lengend Snippet: Molecular docking score matrix for the ten highest‐degree database‐derived candidate compounds and AKT1, STAT3, and RAGE. Values are predicted AutoDock Vina scores in kcal/mol; more negative values indicate more favorable predicted scores within this computational model. The matrix is presented for comparative hypothesis generation and does not establish high‐affinity binding, target engagement, or constituent‐level activity in vivo. Three representative complexes were subsequently evaluated by 100‐ns MD simulations (Figures ).

Article Snippet: Primary antibodies included AGE (A16422, ABclonal; 1:2000), RAGE (66833‐1‐Ig, Proteintech; 1:2000), STAT3 (60199‐1‐Ig, Proteintech; 1:1000), AKT1 (10176‐2‐AP, Proteintech; 1:2000), phospho‐STAT3 (Ser727; AP0715, ABclonal; 1:2000), phospho‐AKT (Ser473; 80455‐1‐RR, Proteintech; 1:1000), and GAPDH (60004‐1‐Ig, Proteintech; 1:20000).

Techniques: Docking Assay, Derivative Assay, Binding Assay, Drug discovery, Activity Assay, In Vivo

Journal: Chemistry & Biodiversity

Article Title: Network Pharmacology and In Vivo Evaluation of Modified Huanglian Wendan Decoction in Polycystic Ovary Syndrome With Insulin Resistance: Association With AGE–RAGE‐Related Signaling

doi: 10.1002/cbdv.71737

Figure Lengend Snippet: Ovarian AGE–RAGE/STAT3/AKT1‐related protein expression and phosphorylation. (A) Representative Western blot bands for AGE, RAGE, STAT3, AKT1, phospho‐STAT3 (Ser727), phospho‐AKT (Ser473), and GAPDH in the Control, Model, HLWDD‐L, HLWDD‐M, HLWDD‐H, and Metformin groups. (B) Relative densitometric analysis of AGE/GAPDH, RAGE/GAPDH, STAT3/GAPDH, AKT1/GAPDH, p‐STAT3/STAT3, and p‐AKT/AKT1. ** p < 0.01 vs. Control; ## p < 0.01 vs. Model. Densitometric comparisons were based on independent biological replicates and analyzed as described in Methods.

Article Snippet: Primary antibodies included AGE (A16422, ABclonal; 1:2000), RAGE (66833‐1‐Ig, Proteintech; 1:2000), STAT3 (60199‐1‐Ig, Proteintech; 1:1000), AKT1 (10176‐2‐AP, Proteintech; 1:2000), phospho‐STAT3 (Ser727; AP0715, ABclonal; 1:2000), phospho‐AKT (Ser473; 80455‐1‐RR, Proteintech; 1:1000), and GAPDH (60004‐1‐Ig, Proteintech; 1:20000).

Techniques: Expressing, Phospho-proteomics, Western Blot, Control, Analysis

Journal: Chemistry & Biodiversity

Article Title: Network Pharmacology and In Vivo Evaluation of Modified Huanglian Wendan Decoction in Polycystic Ovary Syndrome With Insulin Resistance: Association With AGE–RAGE‐Related Signaling

doi: 10.1002/cbdv.71737

Figure Lengend Snippet: Working model integrating the computational and in vivo findings. Network pharmacology and docking prioritized AGE–RAGE/STAT3/AKT1‐related signaling as a candidate network; 100‐ns MD and MM/PBSA analyses supported the dynamic plausibility of three representative docked complexes. In vivo treatment was associated with reproductive, insulin‐related metabolic (including HOMA‐IR), inflammatory, total‐protein, and phosphorylation‐level changes. Dashed connectors denote association; direct target engagement and causal pathway mediation were not established.

Article Snippet: Primary antibodies included AGE (A16422, ABclonal; 1:2000), RAGE (66833‐1‐Ig, Proteintech; 1:2000), STAT3 (60199‐1‐Ig, Proteintech; 1:1000), AKT1 (10176‐2‐AP, Proteintech; 1:2000), phospho‐STAT3 (Ser727; AP0715, ABclonal; 1:2000), phospho‐AKT (Ser473; 80455‐1‐RR, Proteintech; 1:1000), and GAPDH (60004‐1‐Ig, Proteintech; 1:20000).

Techniques: In Vivo, Docking Assay, Phospho-proteomics, Drug discovery