Journal: Cell Reports Medicine

Article Title: The characteristics of pre-existing humoral imprint determine efficacy of S. aureus vaccines and support alternative vaccine approaches

doi: 10.1016/j.xcrm.2023.101360

Figure Lengend Snippet:

Article Snippet: Total mouse IgG were purified from naive and immune mouse sera using Pierce Protein G Agarose (ThermoFisher Scientific), and total human IgG were purified from human sera using Pierce Protein A Agarose (ThermoFisher Scientific).

Techniques: Recombinant, Isolation, Software

Journal: Journal of Cancer

Article Title: FGF10: Type III Epithelial Mesenchymal Transition and Invasion in Breast Cancer Cell Lines

doi: 10.7150/jca.7797

Figure Lengend Snippet: Gene expression and protein modification. a) Quantitative Real time PCR for gene expression analysis in different genes involved in EMT and cell cycle in MCF-7 cells treated by FGF-10 recombinant protein. b) Same analysis in MDA-MB-231 cells treated with FGF-10 siRNA. c) Western Blot analysis has shown the phosphorylation (inactivation) of GSK3β with the same direction of FGF-10 expression in both cell lines, demonstrating that the phosphorylation of GSK3β is increasing when FGF10 is up regulated (in MCF-7, left) and decreasing during FGF-10 down regulation (in MDA-MB-231, right).

Article Snippet: The membranes were rinsed 3 times with TBST and incubated overnight at 4°C with a monoclonal antibody against human FGF-10 (1:1000), a monoclonal anti-human GSK3β (total and p Ser -9) antibody (Santa cruz, USA) and an anti-human actin monoclonal antibody (1:5000) (Santa cruz, USA) in TBST.

Techniques: Gene Expression, Modification, Real-time Polymerase Chain Reaction, Recombinant, Western Blot, Phospho-proteomics, Expressing

Journal: Journal of Cancer

Article Title: FGF10: Type III Epithelial Mesenchymal Transition and Invasion in Breast Cancer Cell Lines

doi: 10.7150/jca.7797

Figure Lengend Snippet: Different mechanisms of signaling for EMT which might be induced by FGF10. FGF10 has a direct effect on its expression, followed by TGF-β up regulation. FGF10 may have inducing effect on EMT through Ras/MAPK and AKT/Pi3K/mTOR pathways. Another finding has shown that WNT signaling activity though GSK3β phosphorylation leads to βCatenin (βCat) localization into the nucleus and Lef/TCF activation. Also GSK3β phosphorylation (inactivation) leads to stabilization of Smad 3 and sensitivity of cells to TGF-β which induces another way of EMT through TGF-β/Smad pathway.

Article Snippet: The membranes were rinsed 3 times with TBST and incubated overnight at 4°C with a monoclonal antibody against human FGF-10 (1:1000), a monoclonal anti-human GSK3β (total and p Ser -9) antibody (Santa cruz, USA) and an anti-human actin monoclonal antibody (1:5000) (Santa cruz, USA) in TBST.

Techniques: Expressing, Activity Assay, Phospho-proteomics, Activation Assay

Journal:

Article Title: Feeding Rapidly Stimulates Protein Synthesis in Skeletal Muscle of Neonatal Pigs by Enhancing Translation Initiation 1–3

doi: 10.3945/jn.109.106781

Figure Lengend Snippet: Time course of the changes in the 4E-BP1·eIF4E complex (A) and eIF4E·eIF4G complex (B) and the phosphorylation of eIF4G (C) in skeletal muscle of neonatal pigs after a meal. All results are corrected for total protein. ANOVA showed an effect of treatment (P < 0.001) for each complex and eIF4G phosphorylation (P < 0.005). Values are means ± SEM, n = 6. Means without a common letter differ, P < 0.05.

Article Snippet: Primary antibodies that were used in the immunoblotting were PKB (total and Ser 473 , Cell Signaling), AMP-activated kinase (AMPK) (total and Thr 172 , Cell Signaling), mTOR (total and Ser 2448 , Cell Signaling), raptor (total and Ser 792 , Cell Signaling), 4E-BP1 (total, Bethyl Laboratories, and Thr 70 , Cell Signaling), eIF4G (total and Ser 1180 , Cell Signaling), S6 (total, Ser 235/236 , and Ser 240/244 , Cell Signaling), eIF2 α (total and Ser 51 , Cell Signaling), and eEF2 (total and Thr 56 , Cell Signaling).

Techniques:

Journal: Pediatric research

Article Title: Insulin modulates energy and substrate sensing and protein catabolism induced by chronic peritonitis in skeletal muscle of neonatal pigs

doi: 10.1038/pr.2016.129

Figure Lengend Snippet: Effect of insulin on the phosphorylation of protein kinase B (p-PKB) (a), tuberous sclerosis complex 2 (p-TSC2) (b), mammalian target of rapamycin (p-mTOR) (c), ribosomal protein S6 kinase-1 (p-S6K1) (d), and the abundance of eukaryotic initiation factor 4 complex (eIF4E-eIF4G) (e) during a 2-h pancreatic glucose–amino acid clamp after 5 d of peritonitis in skeletal muscle of neonatal pigs. CON, control, n = 4; CON + Insulin, n = 5; CLP,cecal ligation and puncture, n = 4; CLP + Insulin, n = 5. Values are means ± SE. * P = 0.05, ** P = 0.001.

Article Snippet: Primary antibodies used in the immunoblotting were against PKB/Akt (total and Ser473; Cell Signaling Technology, Beverly, MA), TSC2 (total and Thr1462; Cell Signaling), mTOR (total and Ser2448; Cell Signaling), S6K1 (total and Thr389; Cell Signaling), AMPKα (total and Thr172; Cell Signaling), atrogin-1 (ECM Biosciences, Versailles, KY), MuRF1 (R&D Systems, Minneapolis, MN), FOXO1 (total and Ser 256; Cell Signaling), FOXO3 (total and Ser253; Cell Signaling), FOXO4 (total and Ser262; Cell Signaling), NF-κB (total and Ser536; Cell Signaling), SIRT1 (Cell Signaling), Pro-caspase 3 (Cell Signaling), and eIF4E-eIF4G (gift of Dr Leonard Jefferson, College of Medicine, Hershey, PA).

Techniques: Ligation

Journal: PLoS ONE

Article Title: The Acute Phase Reactant Orosomucoid-1 Is a Bimodal Regulator of Angiogenesis with Time- and Context-Dependent Inhibitory and Stimulatory Properties

doi: 10.1371/journal.pone.0041387

Figure Lengend Snippet: (A). ORM1 inhibits injury- and TNFα (20 ng/ml)-induced phosphorylation of MEK1/2 and p38, but has no effect on TNFα -induced phosphorylation of NFκB. (B–C) Injury-induced (B) and TNFα-induced (C) aortic angiogenesis are suppressed by the MEK1/2 inhibitor PD98059 (N = 4; *** = p<0.001) whereas inhibitors of other signaling pathways have no effect or are minimally inhibitory (* = p<0.05). (D). ORM1 inhibits TNFα-induced phosphorylation of MEK1/2 and p38, but has no inhibitory effect on TNFα -induced phosphorylation of NFκB in endothelial cells and macrophages; ORM1 has no effect on TNFα signaling in mural cells. (E). ORM1 has no inhibitory effect on VEGF (10 ng/ml) signaling in endothelial cells. (F). ORM1 dose dependently induces phosphorylation of NFκB in endothelial cells and macrophages, but has no significant effect on MEK1/2 and p38 phosphorylation.

Article Snippet: Antibodies against phosphorylated or total MEK1/2 (Ser217/221), p38 MAPK (Thr180/Tyr182), and pNFkB/p65 (Ser536) were purchased from Cell signaling Technology.

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: Cardiac Gq Receptors and Calcineurin Activation Are Not Required for the Hypertrophic Response to Mechanical Left Ventricular Pressure Overload

doi: 10.3389/fcell.2021.639509

Figure Lengend Snippet: NFAT-GATA4 signaling in response to angiotensin II infusion after 48 h. (A) Representative Western blots of NFATc4, phosphorylated glycogen synthase kinase-3 beta (GSK3β) and total GSK3β protein expression in the cytoplasm (upper left panel) and nucleus (upper right panel) . GATA4 blots are shown for the nucleus. (B) Cytoplasmic and nuclear data are normalized with GAPDH and histone, respectively (bottom panels) . All Western blot runs were performed in duplicate or triplicate. Results are presented as the mean value of 3–4 hearts per group ± SEM. * p < 0.05, ** p < 0.01, vs. saline treated controls.

Article Snippet: Primary antibodies included: NFATc4 (1:1500 final dilution; Abcam), phosphorylated (p-) GSK3β (Ser9, 1:1500; Cell Signaling), total GSK3β (1:500; Cell Signaling), GATA4 (1:1000; Santa Cruz Biotechnology), CaMKIIδ (1:1000; Santa Cruz Biotechnology), p-HDAC4 (1:1500; Cell Signaling), total HDAC4 (1:1500; Cell Signaling), MEF2A (1:3000; Cell Signaling).

Techniques: Western Blot, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Cardiac Gq Receptors and Calcineurin Activation Are Not Required for the Hypertrophic Response to Mechanical Left Ventricular Pressure Overload

doi: 10.3389/fcell.2021.639509

Figure Lengend Snippet: NFAT-GATA4 signaling in response to TAC after 48 h. (A) Representative Western blots of NFATc4, phosphorylated glycogen synthase kinase-3 beta (GSK3β) and total GSK3β protein expression in the cytoplasm (upper left panel) and nucleus (upper right panel) . GATA4 blots are shown for the nucleus. (B) Cytoplasmic and nuclear data are normalized with GAPDH and histone, respectively (bottom panels) . All Western blot runs were performed in duplicate or triplicate. Results are presented as the mean value of 3–4 hearts per group ± SEM. None of the statistical comparisons between TAC mice and sham-operated controls were significant.

Article Snippet: Primary antibodies included: NFATc4 (1:1500 final dilution; Abcam), phosphorylated (p-) GSK3β (Ser9, 1:1500; Cell Signaling), total GSK3β (1:500; Cell Signaling), GATA4 (1:1000; Santa Cruz Biotechnology), CaMKIIδ (1:1000; Santa Cruz Biotechnology), p-HDAC4 (1:1500; Cell Signaling), total HDAC4 (1:1500; Cell Signaling), MEF2A (1:3000; Cell Signaling).

Techniques: Western Blot, Expressing