Journal: Metabolites

Article Title: The Cholesterol Metabolite Cholest-5-en-3-One Alleviates Hyperglycemia and Hyperinsulinemia in Obese ( db / db ) Mice

doi: 10.3390/metabo12010026

Figure Lengend Snippet: Luciferase activities in NFκB-Luc/CHO-K1 cells. Cells were treated with or without murine TNFα (10 ng/mL in DMSO), cholesterol (100 µM in DMSO), 5-cholestenone (100 µM choles-5-en-3-one in DMSO), or 1 µM IKK2 inhibitor IV. After 7 h of incubation, NFκB luciferase was detected using the ONE-Step TM Luciferase assay system. Values are expressed as the mean ± standard error ( n = 3 wells). * Significant difference at p < 0.05 compared to DMSO (TNFα-) treatment. † Significant difference at p < 0.05 compared to DMSO (TNFα+) treatment.

Article Snippet: In the NFκB-Luc/CHO-K1 cell line (BPS Bioscience, San Diego, CA, USA), fLUC expression is controlled by the NFκB response element located upstream of the TATA promoter and is suitable for monitoring the activity of the NFκB transcription factor through luminescence readout.

Techniques: Luciferase, Incubation

Journal: Scientific Reports

Article Title: Ligand non-competitive GITR antibody prevents formation of the obligatory signal-triggering GITRL: GITR stoichiometry

doi: 10.1038/s41598-025-32541-6

Figure Lengend Snippet: Discovery and characterization of ligand non-competitive NFκB neutralizing anti-GITR antibody. ( A ) Discovery of ligand competitive and non-competitive antibodies from mice immunization campaign measured by antibody mediated % inhibition of human GITRL binding to recombinant human GITR protein via competition ELISA. ( B ) Recombinant GITRL protein induces NFκB signaling in GITR-NFκB-Luciferase-Jurkat cell line. ( C ) Bivalent ligand non-competitive anti-GITR antibody (hGITR-nc-Ab-#1) neutralizes recombinant GITRL protein induced NFκB signaling. ( D ) Monovalent one-armed ligand non-competitive anti-GITR antibody (hGITR-nc-Ab-#1-OA) also neutralizes recombinant GITRL protein induced NFκB signaling.

Article Snippet: GITRL-GITR mediated NFκB signaling was measured using GITR-NFκB-Luciferase-Jurkat cell line (BPS bioscience).

Techniques: Inhibition, Binding Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Luciferase

Journal: Cell Reports Medicine

Article Title: Synergistic T cell signaling by 41BB and CD28 is optimally achieved by membrane proximal positioning within parallel chimeric antigen receptors

doi: 10.1016/j.xcrm.2021.100457

Figure Lengend Snippet: Investigation of co-stimulation delivered by pCAR-M/34 (A) Carboxyfluorescein N-succinimidyl ester (CFSE)-labeled M-CSFR-specific CAR and pCAR T cells were stimulated for 24 h on T47D or T47D FMS tumor monolayers and then flow sorted prior to RNA extraction. Gene set enrichment analysis (GSEA) demonstrated significant cytokine pathway enrichment in pCAR-M/34 T cells compared with all controls. (B) Enriched cytokine-signaling pathways (false discovery rate [FDR] < 0.25; p < 0.1) in pCAR-M/34 pCAR T cells. p < 0.05 for all listed pathways, unless indicated otherwise. (C) “Blue pink o’gram” heatmap of cytokine gene expression in pCAR-M/34 and control T cell populations following stimulation on T47D FMS tumor monolayers. (D) Engineered Jurkat NF-κB reporter cells were co-cultured with T47D or T47D FMS cells for 5 h. Cell lysates were then analyzed for luciferase activity (mean ± SD, n = 3). Effect of tumor necrosis factor alpha (TNF-α) is shown as positive control. M-CSFR-specific CAR and pCAR T cells were re-stimulated each week as described in Figure 2 C. See Figure S3 for additional data.

Article Snippet: NF-κB luciferase reporter Jurkat , BPS Bioscience (distributed by Tebu-Bio) , Cat# 60651.

Techniques: Labeling, RNA Extraction, Expressing, Cell Culture, Luciferase, Activity Assay, Positive Control

Journal: Cell Reports Medicine

Article Title: Synergistic T cell signaling by 41BB and CD28 is optimally achieved by membrane proximal positioning within parallel chimeric antigen receptors

doi: 10.1016/j.xcrm.2021.100457

Figure Lengend Snippet:

Article Snippet: NF-κB luciferase reporter Jurkat , BPS Bioscience (distributed by Tebu-Bio) , Cat# 60651.

Techniques: Recombinant, Staining, Transfection, Enzyme-linked Immunosorbent Assay, Luciferase, Expressing, Clone Assay, Software

Journal: mAbs

Article Title: Intein mediated high throughput screening for bispecific antibodies

doi: 10.1080/19420862.2020.1731938

Figure Lengend Snippet: Kinetic parameters of reconstituted bsAb compared to parental monovalent oaSEEDbodies and bispecific references. Reconstituted bsAb were compared to their parental monovalent antibodies or bispecific references. Antibodies were captured by anti-human Fc biosensors and subjected to respective antigen binding. Melting temperatures were analyzed by thermal shift assays. (ND, Not defined).

Article Snippet: CD40 activation assays were performed using a CD40 NF-kB Luciferase HEK293 Stable reporter cell line (BPS Bioscience).

Techniques: Binding Assay