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: Animal Nutrition

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

doi: 10.1016/j.aninu.2025.08.011

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

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

Techniques: Binding Assay

Journal: Molecular Medicine Reports

Article Title: RUVBL1 and CRAF promote periodontal ligament stem cell osteogenic differentiation via the MEK/ERK signaling cascade

doi: 10.3892/mmr.2026.13881

Figure Lengend Snippet: Overexpression of CRAF activates the RAF/MEK/ERK signaling pathway. Reverse transcription-quantitative PCR was used to analyze the mRNA expression of (A) ARAF, (B) BRAF, (C) CRAF, (D) MEK1, (E) MEK2, (F) ERK1and (G) ERK2. Western blotting was used to analyze the protein levels of (H) ARAF, (I) BRAF, (J) CRAF, (K) MEK1/2, (L) p-MEK1/2, (M) p-MEK1/2 to MEK1/2, (N) ERK1/2, (O) p-ERK1/2 and (P) p-ERK1/2 to ERK1/2. *P<0.05, **P<0.01 and ***P<0.001. p-, phosphorylated; OE, overexpression; NC, negative control; sh, short hairpin; ns, not significant; ARAF, A-Raf proto-oncogene serine/threonine-protein kinase; BRAF, B-Raf proto-oncogene serine/threonine-protein kinase; CRAF, C-Raf proto-oncogene serine/threonine-protein kinase.

Article Snippet: The antibodies were as follows: GAPDH (1:1,000; cat. no. P30008M; Abmart Pharmaceutical Technology Co., Ltd.), A-Raf proto-oncogene serine/threonine-protein kinase (ARAF) (cat. no. bs-2251R), CRAF (cat. no. bs-23170R), MEK1/2 (cat. no. bs-1041R), phosphorylated (p)-MEK1/2 (cat. no. bs-3270R), ERK1/2 (cat. no. bsm-33232M), p-ERK1/2 (cat. no. bs-3016R; all BIOSS), RUVBL1 (cat. no. 74775; Cell Signaling Technology, Inc.), ERK1/2 (all 1:500; cat. no. bsm-33232M; BIOSS), B-Raf proto-oncogene serine/threonine-protein kinase (BRAF) (1:2,000; cat. no. ab33899; Abcam) and HRP-conjugated universal secondary antibody [cat. nos.

Techniques: Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Negative Control

Journal: Molecular Medicine Reports

Article Title: RUVBL1 and CRAF promote periodontal ligament stem cell osteogenic differentiation via the MEK/ERK signaling cascade

doi: 10.3892/mmr.2026.13881

Figure Lengend Snippet: Overexpression of RUVBL1 activates the MEK/ERK signaling pathway. Reverse transcription-quantitative PCR was used to analyze the mRNA expression of (A) ARAF, (B) BRAF, (C) CRAF, (D) MEK1, (E) MEK2, (F) ERK1 and (G) ERK2. Western blotting was used to analyze protein levels of (H) ARAF, (I) BRAF, (J) CRAF, (K) MEK1/2, (L) p-MEK1/2, (M) p-MEK1/2 to MEK1/2, (N) ERK1/2, (O) p-ERK1/2 and (P) p-ERK1/2 to ERK1/2. *P<0.05, **P<0.01 and ***P<0.001. RUVBL1, RuvB-like AAA ATPase-1; p-, phosphorylated; OE, overexpression; NC, negative control; sh, short hairpin; ns, not significant; ARAF, A-Raf proto-oncogene serine/threonine-protein kinase; BRAF, B-Raf proto-oncogene serine/threonine-protein kinase; CRAF, C-Raf proto-oncogene serine/threonine-protein kinase.

Article Snippet: The antibodies were as follows: GAPDH (1:1,000; cat. no. P30008M; Abmart Pharmaceutical Technology Co., Ltd.), A-Raf proto-oncogene serine/threonine-protein kinase (ARAF) (cat. no. bs-2251R), CRAF (cat. no. bs-23170R), MEK1/2 (cat. no. bs-1041R), phosphorylated (p)-MEK1/2 (cat. no. bs-3270R), ERK1/2 (cat. no. bsm-33232M), p-ERK1/2 (cat. no. bs-3016R; all BIOSS), RUVBL1 (cat. no. 74775; Cell Signaling Technology, Inc.), ERK1/2 (all 1:500; cat. no. bsm-33232M; BIOSS), B-Raf proto-oncogene serine/threonine-protein kinase (BRAF) (1:2,000; cat. no. ab33899; Abcam) and HRP-conjugated universal secondary antibody [cat. nos.

Techniques: Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Negative Control

Journal: Molecular Medicine Reports

Article Title: RUVBL1 and CRAF promote periodontal ligament stem cell osteogenic differentiation via the MEK/ERK signaling cascade

doi: 10.3892/mmr.2026.13881

Figure Lengend Snippet: ODN MT01 promotes osteogenic differentiation of PDLSCs. (A) PDLSC proliferation and (B) ALP staining following treatment with varying concentrations of ODN MT01. (C) Osteogenic differentiation and (D) mineralization after the addition of ODN MT01 and osteogenic inducers. ALP staining was used to assess the effect of (E) CRAF and (F) RUVBL1 on osteogenic differentiation of PDLSCs following the addition of ODN MT01 and osteogenic inducers. Alizarin Red staining was used to assess the effect of (G) CRAF and (H) RUVBL1 on the degree of mineralization of PDLSCs following the addition of ODN MT01 and osteogenic inducers. *P<0.05 and ***P<0.001. ODN, oligodeoxynucleotide; PDLSC, periodontal ligament stem cell; ALP, alkaline phosphatase, RUVBL1, RuvB-like AAA ATPase-1; NC, negative control; OE, overexpression; sh, short hairpin; ns, not significant; CRAF, C-Raf proto-oncogene serine/threonine-protein kinase; OI, osteogenic induction; OM, ODN MT01.

Article Snippet: The antibodies were as follows: GAPDH (1:1,000; cat. no. P30008M; Abmart Pharmaceutical Technology Co., Ltd.), A-Raf proto-oncogene serine/threonine-protein kinase (ARAF) (cat. no. bs-2251R), CRAF (cat. no. bs-23170R), MEK1/2 (cat. no. bs-1041R), phosphorylated (p)-MEK1/2 (cat. no. bs-3270R), ERK1/2 (cat. no. bsm-33232M), p-ERK1/2 (cat. no. bs-3016R; all BIOSS), RUVBL1 (cat. no. 74775; Cell Signaling Technology, Inc.), ERK1/2 (all 1:500; cat. no. bsm-33232M; BIOSS), B-Raf proto-oncogene serine/threonine-protein kinase (BRAF) (1:2,000; cat. no. ab33899; Abcam) and HRP-conjugated universal secondary antibody [cat. nos.

Techniques: Staining, Negative Control, Over Expression