primary antibody against p ir β  (Cell Signaling Technology Inc)

Journal: Cell Discovery

Article Title: A decrease in Flavonifractor plautii and its product, phytosphingosine, predisposes individuals with phlegm-dampness constitution to metabolic disorders

doi: 10.1038/s41421-025-00789-x

Figure Lengend Snippet: Key materials and software.

Article Snippet: Primary antibody against p-IR-β , Cell Signaling Technology, Inc., Shanghai, China , #2381.

Techniques: Software, Recombinant, Luciferase, Reporter Gene Assay, Plasmid Preparation, Control, Transfection, Enzyme-linked Immunosorbent Assay, Staining

Journal: bioRxiv

Article Title: Follicular mural granulosa cells stockpile glycogen to fuel corpus luteum pre-vascularization

doi: 10.1101/2025.01.22.634063

Figure Lengend Snippet: (A) KEGG analysis revealed activated signaling in fGCs post-ovulation/luteinization. (B) Western blotting displayed alterations in insulin receptor activity in fGCs post-stimulation. (C-E) Insulin signaling mediates hCG-induced glucose uptake and glycogen storage in fGCs. (C) Protein levels related to glucose uptake, glycogen synthesis, and glycogenolysis in fGCs were influenced by activation or inhibition of insulin signaling. (D) Flow cytometry measured changes in fGC glucose uptake capacity upon activation or inhibition of insulin signaling. Left: Representative images. Right: Statistical chart; n = 4 independent fGC samples. (E) Impact of activating or inhibiting insulin signaling on fGC glycogen content; scale bar = 100 μm. (F) Bubble plot of the top 10 transcription factors significantly induced by hCG in fGCs. (G) ChIP-seq analysis revealed RUNX1 binding to Insr and Igf1r promoters. Arrows indicate RUNX1 binding peaks. (H) Effects of inhibiting or activating the Ras/Raf/Mek/Erk signaling cascade on RUNX1, glucose uptake-related, and glycogen synthesis-related protein levels in fGCs. (I) PAS staining illustrated the influence of inhibiting or activating Ras/Raf/Mek/Erk on fGC glycogen content; scale bar = 100 μm. (J) EMSA demonstrated RUNX1 binding to Insr and Igf1r promoter sequences. (K) ChIP-qPCR assay for RUNX1 binding to Insr and Igf1r promoters. Input and IgG are positive and negative controls, respectively. UP: qPCR statistical chart; n = 3 independent fGC samples. (L) Dual-luciferase reporter assay identified specific motifs within Insr and Igf1r promoters interacting with RUNX1; n = 3 independent samples. (M) Knockdown of RUNX1 impacted glycogen synthesis-related protein levels and glycogen content in fGCs. Left: Experimental design. Right: Western blotting and PAS staining; scale bar = 100 μm. Statistical significance were determined using one-way ANOVA followed by Tukey’s post hoc test, values were mean ± SD. Significant differences were denoted by *P<0.05, **P<0.01, ****P<0.001,****P<0.0001.

Article Snippet: After transfer, the membrane was blocked with 5% skim milk powder (Nestle, Switzerland) at room temperature, followed by overnight incubation at 4°C with specific primary antibodies: P-PKCα (1:1000 dilution; 9375T, CST, USA), SLC2A1 (1:1000 dilution; 21829-1-AP, Proteintech, USA), GYS1 (1:1000 dilution; 3886T, CST, USA), GSK3B (1:1000 dilution; 12456T, CST, USA), P-GYS1 (1:1000 dilution; 47043T, CST, USA), P-GSK3B (1:1000 dilution; 9323T, CST, USA), PYGB (1:1000 dilution; 12075-1-AP, Proteintech, USA), UGP2 (1:800 dilution; 10391-1-AP, Proteintech, USA), INSR/IGF1R (1:1000 dilution; A21984, Abclonal, China), P-INSR/IGF1R (1:800 dilution; 3024T, CST, USA), RUNX1 (1:1000 dilution; 25315-1-AP, Proteintech, USA), ERK1/2 (1:1000 dilution; A4782, Abclonal, China), P-ERK1/2 (1:1000 dilution; AP0234, Abclonal, China), GAPDH (1:5000 dilution; AC002, Abclonal, China).

Techniques: Western Blot, Activity Assay, Activation Assay, Inhibition, Flow Cytometry, ChIP-sequencing, Binding Assay, Staining, Luciferase, Reporter Assay, Knockdown

Journal: Journal of Translational Medicine

Article Title: VEGFB ameliorates insulin resistance in NAFLD via the PI3K/AKT signal pathway

doi: 10.1186/s12967-024-05621-w

Figure Lengend Snippet: Antibodies (human and mouse) used in this study

Article Snippet: p-INSR Thr1375 , 1:1000 , Rabbit , Abmart.

Techniques:

Journal: Journal of Translational Medicine

Article Title: VEGFB ameliorates insulin resistance in NAFLD via the PI3K/AKT signal pathway

doi: 10.1186/s12967-024-05621-w

Figure Lengend Snippet: VEGFB activates PI3K/AKT signal pathway in insulin-resistant HepG2 cells ( A ) Intracellular glucose content ( n = 3) ( B ) Intracellular glycogen content ( n = 3) ( C ) PPI analysis ( D ) VEGFR1, INSR, p-INSR Thr1375 , IRS1, p-IRS1 Ser307 , PI3K, p-PI3K Tyr467 , AKT, p-AKT Ser473 , GLUT2, GSK3β, p-GSK3β Ser9 , FOXO1, and p-FOXO1 pS256 proteins expression were detected by western blot ( E ) VEGFR1/β-actin expression ( n = 3) ( F ) p-INSR Thr1375 /INSR expression ( n = 3) ( G ) p-IRS1 Ser307 /IRS1 expression ( n = 3) ( H ) p-PI3K Tyr467 /PI3K expression ( n = 3) ( I ) p-AKT Ser473 /AKT expression ( n = 3) ( J ) GLUT2/β-actin expression ( n = 3) ( K ) p-GSK3β Ser9 /GSK3β expression ( n = 3) ( L ) p-FOXO1 pS256 /FOXO1 expression ( n = 3) ( M ) Effects of VEGFB on VEGFR1, INSR, IRS1, PI3K, AKT, GLUT2, G6Pase, and PEPCK genes expression at mRNA level ( n = 3) * indicates to compare with HepG2 group, # indicates to compare with IR + siRNA-NC group, Δ indicates to compare with IR + LV-NC group with a significant difference p < 0.05

Article Snippet: p-INSR Thr1375 , 1:1000 , Rabbit , Abmart.

Techniques: Expressing, Western Blot

Journal: Journal of Translational Medicine

Article Title: VEGFB ameliorates insulin resistance in NAFLD via the PI3K/AKT signal pathway

doi: 10.1186/s12967-024-05621-w

Figure Lengend Snippet: Effects of VEGFB on hepatic insulin resistance via PI3K/AKT signal pathway ( A ) VEGFR1, INSR, p-INSR Thr1375 , IRS1, p-IRS1 Ser307 , PI3K, p-PI3K Tyr467 , AKT, p-AKT Ser473 , GLUT2, GSK3β, p-GSK3β Ser9 , FOXO1, and p-FOXO1 pS256 proteins expression ( B ) VEGFR1/β-actin expression ( C ) p-INSR Thr1375 /INSR expression ( D ) p-IRS1 Ser307 /IRS1 expression ( E ) p-PI3K Tyr467 /PI3K expression ( F ) p-AKT Ser473 /AKT expression ( G ) GLUT2/β-actin expression ( H ) p-GSK3β Ser9 /GSK3β expression ( I ) p-FOXO1 pS256 /FOXO1 expression ( J ) Effects of VEGFB on VEGFR1, IRS1, PI3K, AKT, GLUT2, INSR, PEPCK, and G6Pase genes expression at mRNA level ( K ) Hepatic glucose content ( L ) Hepatic glycogen content * indicates to compare with SD group, # indicates to compare with HFD-WT group, Δ indicates to compare with HFD-KO group with a significant difference p < 0.05

Article Snippet: p-INSR Thr1375 , 1:1000 , Rabbit , Abmart.

Techniques: Expressing

Journal: The FASEB Journal

Article Title: Cysteine‐rich 61 inhibition attenuates hepatic insulin resistance and improves lipid metabolism in high‐fat diet fed mice and HepG2 cells

doi: 10.1096/fj.202400860R

Figure Lengend Snippet: List of antibodies used for western blotting.

Article Snippet: Antibodies against Cyr61 (#39382), Sirt6 (#12486), phosphorylated (p)‐AMPKα (Thr172) (#2531), AMPKα (#2532), p‐insulin receptor (IR) β (Tyr1361) (#3023), IR β (#3025), insulin receptor substrate 1 (IRS‐1) (#2382), p‐ protein kinase B (Akt) (Ser473) (#9271), Akt (#9272), p‐ glycogen synthase kinase‐3 alpha/beta (GSK‐3α/β) (Ser21/9) (#8566), GSK‐3α/β (#5676), p‐ nuclear factor kappa B (NF‐κB) p65 (Ser536) (#3033), NF‐κB p65 (#4764), p‐stress‐activated protein kinase (SAPK)/ c‐Jun N‐terminal kinase (JNK) (Thr183/Tyr185) (#9251), SAPK/JNK (#9252), and α‐tubulin (#2144) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Western Blot