nfat luciferase reporter jurkat cells  (BPS Bioscience)

Journal: Toxicological Sciences

Article Title: Nonclinical safety assessment of SBT777101, a CAR-engineered regulatory T cell therapy for autoimmune diseases

doi: 10.1093/toxsci/kfag125

Figure Lengend Snippet: Selective activation of SBT777101 Jurkat reporter cells by citrullinated proteins. Representative dose–response curves showing fold-increase in luminescence ( y -axis) relative to baseline following incubation with increasing concentrations (log scale, x -axis) of citrullinated (red) or noncitrullinated (black) protein targets. Data points represent the mean ± SD of 3 replicates.

Article Snippet: NFAT-luciferase reporter Jurkat cells (BPS Bioscience) were stably transduced with the SBT777101 CAR.

Techniques: Activation Assay, Incubation

Journal: Nature

Article Title: GPCR antagonism via rewiring of receptor trafficking and degradation

doi: 10.1038/s41586-026-11088-0

Figure Lengend Snippet: (a) Schematic of native versus engineered GPCR trafficking (left). Native agonists typically induce GPCR internalization, followed by recycling or degradation. GTACs direct GPCRs for degradation by leveraging rapid, constitutive TfR1-mediated internalization. GTAC design (right): bifunctional antibody with GPCR-binding domain(s) and TfR1-binding domain(s) fused to an Fc. The control antibody lacks TfR1-binding domain(s). (b) Cell-surface GPCR levels (anti-FLAG flow cytometry) after treatment with GTACs or control antibodies. Cell-surface AT1R levels in FLAG-AT1R-GFP + TfR1 + Expi293 cells after 6 h treatment; cell-surface RXFP1 levels in FLAG-RXFP1 + TfR1 + Expi293 cells after 2 h treatment; cell-surface APJ levels in FLAG-APJ + TfR1 + HEK293T cells after 2 h treatment; and cell-surface A2AR levels in FLAG-A2AR + TfR1 + Expi293 cells after 6 h treatment. Data represent mean ± standard deviation (s.d.) from n = 3 replicates. (c) NFAT-luciferase activation in FLAG-AT1R + TfR1 + NFAT-Luc + Expi293 cells, after 2 h pre-treatment with 50 nM (left) or varying concentration (right) AT1R GTAC or Nb-Fc, followed by 4 h stimulation with varying concentration (left) or 9 nM (right) AngII. Data represent mean ± s.d. from n = 4 biological replicates. (d) β-arrestin recruitment in FLAG-AT1R-Tango + TfR1 + HTLA cells (luminescence) after pre-treatment for 2 h with 50 nM (left) or varying concentration (right) AT1R GTAC or Nb-Fc, followed by 14 h activation with varying concentrations (left) or 200 nM (right) AngII. Data represent mean ± s.d. from n = 3 biological replicates for both experiments. (e) RLN2-induced cAMP signaling in FLAG-RXFP1 + TfR1 + GloSensor + HEK293T cells (luminescence) after 2 h pre-treatment with 10 nM (left) or varying concentration (right) RXFP1 GTAC or control, followed by 30 min stimulation with RLN2 at varying concentrations (left) or 1 nM (right). Data represent mean ± s.d. from n = 3 biological replicates.

Article Snippet: NFAT luciferase reporter was a gift from Toren Finkel (Addgene plasmid #10959; http://n2t.net/addgene:10959 ; RRID:Addgene_10959).

Techniques: Binding Assay, Control, Flow Cytometry, Standard Deviation, Luciferase, Activation Assay, Concentration Assay

Journal: Cells

Article Title: NeoPAIR-T: Functional Mapping of Neoantigen–TCR Pairs Using a CRISPR-Engineered Jurkat Reporter System

doi: 10.3390/cells14221789

Figure Lengend Snippet: Establishment of reporter T cells and CRISPR/Cas9-mediated knock-in of a candidate T cell receptor (TCR). ( A ) Surface marker expression of parental Jurkat E6-1 cells. Histograms show the expression levels of the indicated markers. ( B ) Surface marker expression of engineered reporter T cells. Histograms show the expression levels of the indicated markers. ( C ) Schematic overview of the genetic modifications performed to generate reporter T cells. Parental Jurkat E6-1 cells were subjected to CRISPR/Cas9-mediated knockout of CD4, FAS, and TCRα, and lentiviral transduction of CD8α, CD8β, Cas9, and the NFAT–Luc–P2A–eGFP reporter cassette. ( D ) Targeted reconstitution of the TCR in reporter T cells at the endogenous TCRβ locus. ( E ) Flow cytometric analysis of CD3 restoration in reporter T cells after targeted reconstitution with the NY-ESO-1–specific 1G4 TCR. CD3 + cells were sorted to generate a uniform population of TCR-expressing reporter T cells. Pseudocolor plots indicate event density (blue, lower; red, higher).

Article Snippet: Finally, these cells were transduced with a lentiviral NFAT-Luciferase-eGFP reporter construct ( , BPS Bioscience, San Diego, CA, USA) at MOI of 20 and selected with 2 mg/mL G418 (FujiFilm Wako Pure Chemical Corporation).

Techniques: CRISPR, Knock-In, Marker, Expressing, Knock-Out, Transduction