anti phosphorylated p insulin receptor substrate 1  (Cell Signaling Technology Inc)

Journal: eNeuro

Article Title: AD-Like Neuropsychiatric Dysfunction in a Mice Model Induced by a Combination of High-Fat Diet and Intraperitoneal Injection of Streptozotocin

doi: 10.1523/ENEURO.0310-24.2024

Figure Lengend Snippet: Combination of HFD and STZ/I.P. induced the dysregulation of the IRS1/AKT/ERK signaling pathway in mice. A , The typical graph of IRS1/AKT/ERK signaling pathway proteins in the hippocampus and PFC of mice. B , The statistical analysis of the Western blotting results in the hippocampus. C , The statistical analysis of the Western blotting results in the PFC. The data are presented as the mean ± SEM, with n = 3 in each group. * p < 0.05 and ** p < 0.01 compared with the Con group.

Article Snippet: Equal amounts of protein were separated on 10% SDS–PAGE gels, transferred onto polyvinylidene difluoride (PVDF) membranes, and incubated overnight at 4°C with the relevant primary antibodies: anti-phosphorylated AKT (p-AKT) and total AKT (1:1,000; Zen-Bio), anti-phosphorylated ERK (p-ERK) and total ERK (1:1,000; Santa Cruz Biotechnology), anti-phosphorylated IRS1 (p-IRS1) and total IRS1 (1:1,000; Abcam), anti-APP(1:1,000; Proteintech), anti-phosphorylated Tau (p-Tau) and total tau (1:1,000; Santa Cruz Biotechnology), anti-TREM1 and TREM2 (1:1,000; Abcam), and anti-β-actin (1:1,000; Zhongshan Biotechnology).

Techniques: Western Blot

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Blautia Coccoides is a Newly Identified Bacterium Increased by Leucine Deprivation and has a Novel Function in Improving Metabolic Disorders.

doi: 10.1002/advs.202309255

Figure Lengend Snippet: Figure 5. I3AA has beneficial effects on metabolic improvements in vitro and in vivo. A) Primary hepatocytes were incubated with indicate dose of I3AA for 48 h and then stimulated with 100 nM insulin for 20 min (n = 6 replicates per group). Western blot analysis of p-IR, p-AKT, and p-GSK3𝛽levels. The right panel is the densitometry analysis of the relative abundance of phosphorylated proteins normalized to their total protein levels. A.U.: arbitrary units. B–J) HFD mice were orally gavage with PBS or 10 mg kg−1 I3AA for 4 weeks (n = 6–7 biological replicates per group). B) Fed and fasting blood glucose levels. C) Fed and fasting serum insulin levels assayed by ELISA. D) HOMA-IR index. E) Glucose tolerance tests. The right panel is the AUC. F) Insulin tolerance tests (0.5 U kg−1). The right panel is the AUC. G) Total fat mass. H) The H&E staining of sWAT. Scale bars, 50 μm. The bottom panel is the frequency distribution of adipocyte cell size in sWAT and the box plot is average adipocyte diameter. I) Real-time PCR analysis of browning related genes (Ucp1, Pgc1𝛼, Prdm16, and Cidea) in sWAT. J) Western blot analysis of UCP1 protein levels. The bottom panel is the densitometry analysis of UCP1 protein levels. All values are expressed as the mean ± SEM. Statistical comparisons were carried out by unpaired two-tailed Student’s t-test; *p < 0.05 and **p < 0.01.

Article Snippet: Protein samples were subjected to concentration determination and immunoblot assay with the following antibodies: phosphorylated (p)-IR (Tyr 1150/1151; 3024S), total IR (3025), p-AKT (Ser 473; 9271S), total AKT (9272S), p-GSK3β (Ser 9; 9336S), and total GSK3β(9315S) from Cell Signaling Technology; AhR (17840-1-AP) and β–Actin (81115-1-RR) from Proteintech; FGF21 (ab171941) and UCP1 (ab209483) from Abcam.

Techniques: In Vitro, In Vivo, Incubation, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Real-time Polymerase Chain Reaction, Two Tailed Test

Journal: Plants

Article Title: Glycolipids Derived from the Korean Endemic Plant Aruncus aethusifolius Inducing Glucose Uptake in Mouse Skeletal Muscle C2C12 Cells

doi: 10.3390/plants13050608

Figure Lengend Snippet: Effects of compounds 1 and 2 on the protein expression levels of phospho-insulin receptor substrate-1 (P-IRS-1), IRS-1, phosphorylated-5′-AMP-activated protein kinase (p-AMPK), AMPK, peroxisome proliferator-activated receptor γ (PPARγ), and glucose transporter type 4 (GLUT4). ( A ) p -IRS-1, IRS-1, p -AMPK, AMPK, PPARγ, GLUT4, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in C2C12 cells treated or untreated with 25 and 50 μM of compounds 1 and 2 . ( B ) Each bar graph presents the densitometric quantification of Western blot bands. The data represent the mean ± S.E.M., n = 3. * p < 0.05 compared to the control.

Article Snippet: The primary antibodies for IRS-1 and GLUT4 and secondary antibodies for phosphorylated-IRS-1 (p-IRS-1) and AMPK, phosphorylated-AMPK (p-AMPK), and PPAR-γ were purchased from Cell Signaling (Danvers, MA, USA).

Techniques: Expressing, Western Blot, Control

Journal: Plants

Article Title: Glycolipids Derived from the Korean Endemic Plant Aruncus aethusifolius Inducing Glucose Uptake in Mouse Skeletal Muscle C2C12 Cells

doi: 10.3390/plants13050608

Figure Lengend Snippet: Docking model for IRS-1 (PDB ID:1QQG). The receptor and ligand are shown in cartoon (cyan color) and stick (magenta color), respectively.

Article Snippet: The primary antibodies for IRS-1 and GLUT4 and secondary antibodies for phosphorylated-IRS-1 (p-IRS-1) and AMPK, phosphorylated-AMPK (p-AMPK), and PPAR-γ were purchased from Cell Signaling (Danvers, MA, USA).

Techniques: