elisa  (Cell Signaling Technology Inc)

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: Tumour Virus Research

Article Title: Oncolytic virus hijacks GOT1 and pyrimidinosomes to fuel pyrimidine synthesis for replication in tumor cells

doi: 10.1016/j.tvr.2026.200342

Figure Lengend Snippet: | The MAS-transported NAD + /NADH regulates CAD phosphorylation activity and NDV replication through S6K. (A) Schematic diagram showing the potential involvement of mTOR-S6K in regulating CAD phosphorylation through MAS-mediated NAD + /NADH translocation. (B-D) A549 cells were either mock-infected or infected with NDV (MOI = 1) at 6, 12, 18 and 24 hpi. Protein levels of p-S6K (T389), S6K, p-S6 (S235/236), S6, p-CAD (S1859), CAD, NP and β-actin were analyzed by WB at 12 hpi (B). Grayscale quantification of p-S6K/S6K, p-S6/S6 and p-CAD/CAD ratios from WB data (C). Intracellular NAD + /NADH ratio were measured (D). (E-H) A549 cells were infected with NDV (MOI = 1), treated with AOA, supplemented with Pyr or Asp. Protein levels of p-S6K, S6K, p-S6, S6, p-CAD, CAD, NP, and β-actin were analyzed by WB at 12 hpi (E). Grayscale quantification of p-S6K/S6K, p-S6/S6 and p-CAD/CAD ratios from WB data (F). Intracellular NAD + /NADH ratio were measured (G). Extracellular virus yields were detected (H). (I-L) A549 cells were infected with NDV (MOI = 1), treated with AOA, supplemented with Pyr, and exposed to Torin 1. Protein levels of p-S6K, S6K, p-S6, S6, p-CAD, CAD, NP, and β-actin were analyzed by WB at 12 hpi (I). Grayscale quantification of p-S6K/S6K, p-S6/S6 and p-CAD/CAD ratios from WB data (J). Intracellular NAD + /NADH ratio were measured (K). Extracellular virus yields were detected (L). (M − P) A549 cells were infected with NDV (MOI = 1), treated with AOA, and supplemented with Duro. Protein levels of p-S6K, S6K, p-S6, S6, p-CAD, CAD, NP, and β-actin were analyzed by WB at 12 hpi (M). Grayscale quantification of p-S6K/S6K, p-S6/S6 and p-CAD/CAD ratios from WB data (N). Intracellular NAD + /NADH ratio were measured (O). Extracellular virus yields were detected (P). Data are presented as means from three independent experiments, ns, p > 0.05, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The antibodies used for immunoblotting in this study included monoclonal or polyclonal: anti-β-actin (Proteintech, Cat# 66009-1-Ig), anti-NP (Prepared in our laboratory), anti-S6 ribosomal protein (Cell Signaling Technology [CST], Cat# 2217), anti-phospho-S6 ribosomal protein (S235/236) (CST, Cat# 4858), anti-phospho-p70 S6 Kinase 1 (T389) (ABclonal, Cat# AP0564), anti-p70 S6 Kinase 1 (ABclonal, Cat# A2190), anti-CAD (CST, Cat# 11933), anti-phospho-CAD (S1859) (CST, Cat# 70307), anti-GOT1 (Proteintech, Cat# 14886-1-AP), anti-GOT2 (Proteintech, Cat# 14800-1-AP). anti-UMPS (Proteintech, Cat# 14830-1-AP), anti-DHODH (Proteintech, Cat# 14877-1-AP) and anti-Tom20 (Abcam, Cat# ab283317).

Techniques: Phospho-proteomics, Activity Assay, Translocation Assay, Infection, Virus