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: 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: Materials Today Bio

Article Title: Extracellular biogenic nanoscale mitochondria reprogram the wound microenvironment via ROS scavenging independent of cellular uptake

doi: 10.1016/j.mtbio.2026.103023

Figure Lengend Snippet: MSC-mt mitigate oxidative damage and improve mitochondrial function in H 2 O 2 -treated skin fibroblasts. (A–B) Representative flow cytometry plots and quantification of Annexin V/PI staining for apoptosis after H 2 O 2 stimulation, showing that MSC-mt significantly reduce fibroblast apoptosis, whereas rotenone-pretreated mitochondria (mt(R)) exhibit markedly attenuated protective effects. (C–D) Representative plots and quantification of mitoSOX staining for mitochondrial ROS after 180 min H 2 O 2 exposure, showing that MSC-mt suppress mitochondrial ROS accumulation, an effect largely lost following rotenone pretreatment. (E–F) Representative plots and quantification of TMRE staining for mitochondrial membrane potential (ΔΨm) after 180 min H 2 O 2 treatment, showing preservation of mitochondrial membrane potential by MSC-mt but not by mt(R). (G–H) Representative plots and quantification of SA-β-gal staining for cellular senescence after 180 min H 2 O 2 treatment, showing reduced senescence burden in MSC-mt–treated cells, with diminished efficacy observed in the mt(R) group. (I) Representative images and quantification of colony formation assays on day 8 post-treatment, indicating improved long-term proliferative capacity following MSC-mt treatment, an effect substantially weakened following rotenone pretreatment. (J) Heatmap of differentially expressed genes identified by a wound healing PCR array following 180 min of H 2 O 2 stimulation, showing that MSC-mt partially reverse H 2 O 2 -induced transcriptional alterations associated with stress response and repair pathways, an effect attenuated when using mt(R). (K) Representative images of mouse apoptosis protein array analysis after 180 min of H 2 O 2 treatment, showing that MSC-mt partially restore the expression of key anti-apoptotic and pro-survival signaling proteins suppressed by oxidative stress, including phosphorylated AKT (Ser473), BAD (Ser112), ERK (T202), and IκBα (S32), with reduced restoration observed in the mt(R) group. (L) Western blot analysis of phosphorylated AKT at Ser473 expression after 180 min of H 2 O 2 exposure, showing restoration of pro-survival signaling by MSC-mt, which is attenuated in the mt(R) group. All experiments were independently repeated three times (n = 3). Data are presented as mean ± SEM. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.

Article Snippet: After blocking with 5% non-fat milk, membranes were incubated overnight at 4 °C with the following primary antibodies: anti-PINK1 (CST, Cat# 6946), anti-NIX (CST, Cat# 12396), anti-TOM20 (CST, Cat# 42406), anti-USP30 (Proteintech, Cat# 15402-1-AP), Total OXPHOS Rodent WB Antibody Cocktail (Abcam, Cat# ab110413), anti-phospho-AKT (Ser473) (CST, Cat# 4060), and anti-β-actin (CST, Cat# 4967).

Techniques: Flow Cytometry, Staining, Membrane, Preserving, Protein Array, Expressing, Western Blot

Journal: International Dental Journal

Article Title: Endothelial Nitric Oxide Synthase Restores Diabetic Dentin Regeneration via AKT/Endothelial Nitric Oxide Synthase Axis

doi: 10.1016/j.identj.2026.109511

Figure Lengend Snippet: Both p-AKT/AKT and eNOS expression are downregulated in dental pulp from diabetic patients. (A and B) Immunohistochemistry staining results of eNOS in dental pulp tissue in NT group and DM group. (C-E) Western Blot confirmed the phosphorylation degree of AKT and eNOS in NT group and DM group. Scale bar: 100 μm. Data presented as mean ± SD.

Article Snippet: After blocking with 5% BSA, the membranes were incubated overnight at 4°C with primary antibodies against: DSPP (1:2000, bs-10316R, Bioss), DMP1 (1:1000, 12359R, Bioss), p-AKT (1:2000, 4060S, CST), AKT (1:500, 60203-2-Ig, Proteintech), eNOS (1:500, ab76198, Abcam), and GAPDH (1:5000, AC026, Proteintech).

Techniques: Expressing, Immunohistochemistry, Staining, Western Blot, Phospho-proteomics

Journal: International Dental Journal

Article Title: Endothelial Nitric Oxide Synthase Restores Diabetic Dentin Regeneration via AKT/Endothelial Nitric Oxide Synthase Axis

doi: 10.1016/j.identj.2026.109511

Figure Lengend Snippet: High glucose affects the proliferation and differentiation of hDPSCs and inhibits the expression of eNOS in hDPSCs via the AKT pathway. (A) Analysis of CCK-8 assay in DPSCs cultured in media with different glucose concentrations. (B) Quantification of ALP activity in DPSCs cultured in media with different glucose concentrations. (C) Quantification of ALP activity in DPSCs from blank, mannitol and HG groups. (D-F) Western Blot confirmed the proteins level of p-AKT, AKT and eNOS in blank, HG and SC79 group. Data are presented as mean ± SD.

Article Snippet: After blocking with 5% BSA, the membranes were incubated overnight at 4°C with primary antibodies against: DSPP (1:2000, bs-10316R, Bioss), DMP1 (1:1000, 12359R, Bioss), p-AKT (1:2000, 4060S, CST), AKT (1:500, 60203-2-Ig, Proteintech), eNOS (1:500, ab76198, Abcam), and GAPDH (1:5000, AC026, Proteintech).

Techniques: Expressing, CCK-8 Assay, Cell Culture, Activity Assay, Western Blot