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 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: GAPDH , Proteintech , 60004-1-Ig.

Techniques: Cell-Signaling

Journal: Pigment Cell & Melanoma Research

Article Title: Allele‐Specific GNAQ Q209 Mutations Reveal Context‐Dependent Oncogenic Programs in Uveal Melanoma

doi: 10.1111/pcmr.70121

Figure Lengend Snippet: Q209R preferentially engages FAK‐YAP signaling. (A) Western blot analysis of GNAQ, BAP1, Melan‐A, p‐ERK1/2 (Thr202/Tyr204), ERK1/2, FRA1, p‐AKT (Ser473), AKT, p‐4EBP1 (Ser65), and 4EBP1 expression in mouse UM cell lines harboring GNAQ Q209R or GNAQ Q209L . HSP90 was used as an internal loading control. (B) Quantification of the relative ratio between p‐AKT (Ser473) and AKT in Q209R and Q209L UM cell lines from (A). (C) Quantification of the relative ratio between p‐4EBP1 (Ser65) and 4EBP1 in Q209R and Q209L UM cell lines from (A). (D) Western blot analysis of p‐FAK (Tyr397), FAK, p‐YAP (Ser127), and YAP in mouse UM cell lines harboring GNAQ Q209R or GNAQ Q209L . HSP90 was used as an internal loading control. (E) Quantification of the relative ratio between p‐FAK (Tyr397) and FAK in Q209R and Q209L UM cell lines from (D). (F) Quantification of the relative ratio between p‐YAP (Ser127) and YAP in Q209R and Q209L UM cell lines from (D). Orthotopic tumor‐derived cell lines (QBM12.1 [Q209R], QBM12.3 [Q209R], QBM24 [Q209R], QBM45 [Q209L], QBM47 [Q209L]) and liver metastatic lines (QBM12L [Q209R], QBM12L2 [Q209R], QBM51L [Q209L], QBM53L [Q209L]) were used in A–F. (G) Representative IHC images showing YAP expression in primary tumors from Gnaq CA ; Bap1 CKO ; Myc CKI mice harboring either GNAQ Q209R or GNAQ Q209L . (H) Quantification of the nuclear‐to‐cytoplasmic YAP intensity ratio (H‐score) among primary tumors from Gnaq CA ; Bap1 CKO ; Myc CKI mice harboring either GNAQ Q209R or GNAQ Q209L . (I) Representative IHC images showing YAP expression in liver metastasis derived from intrasplenic transplantation of mouse UM cell lines harboring GNAQ Q209R or GNAQ Q209L . (J) Quantification of nuclear YAP intensity (H‐score) among liver metastasis derived from intrasplenic transplantation of mouse UM cell lines harboring GNAQ Q209R or GNAQ Q209L .

Article Snippet: Antibodies against GNAQ (CST, #14373, RRID:AB_2665457), BAP1 (Abcam, ab255611), Melan‐A (Abcam, ab210546, RRID:AB_2889292), ERK1/2 (CST, #4695, RRID:AB_390779), p‐ERK1/2 (Thr202/Tyr204) (CST, #4370, RRID:AB_2315112), FRA1 (Abcam, ab252421, RRID:AB_2904513), AKT (CST, #4691, RRID:AB_915783), p‐AKT (Ser473) (CST, #4060, RRID:AB_2315049), p‐FAK (Tyr397) (CST, #8556, RRID:AB_10891442), FAK (CST, #3285, RRID:AB_2269034), 4E‐BP1 (CST, #9452, RRID:AB_331692), p‐4E‐BP1 (Ser65; CST, #9451, RRID:AB_330947), YAP (CST, #4912, RRID:AB_2218911), and p‐YAP (Ser127) (CST, #4911, RRID:AB_2218913) were used at 1:1000 as indicated.

Techniques: Western Blot, Analysis, Expressing, Control, Derivative Assay, Transplantation Assay