Journal: Advanced Science

Article Title: Repurposing a Small Molecule Plant Hormone as a Tunable ON‐Switch for CAR‐T Cell Immunotherapy

doi: 10.1002/advs.77020

Figure Lengend Snippet: Assessment of auxCAR‐T cell performance in Jurkat T cells. (A,B) Representative Flow cytometry (A) and Statistical quantification (B) of early T cell activation as reflected by upregulated CD69 expression on the cell surface. Jurkat T cells transduced with stCAR (left) and auxCAR (right) were co‐cultured with hCD19 + Raji cells or hCD19 − K562 cells, with or without auxin. n = 3 independent biological replicates (mean ± s.d.). The p ‐values were calculated using one‐way analysis of variance (ANOVA). (C,D) Statistical quantification of NFAT‐GFP reporter activity in Jurkat T cells expressing stCAR and auxCAR co‐cultured with tumor cells with hCD19 + Raji cells (C) or A431‐CD19 (D) with the indicated auxin concentration. n = 3 independent technical replicates (mean ± s.d.). The p ‐values were calculated using one‐way analysis of variance (ANOVA). (E) Auxin‐dose‐dependent responses of stCAR T and auxCAR‐T cells examined by IL‐2 production in Jurkat T cells. (F) Architecture of the auxCAR HER2. Upper: cartoon depicting the different components. Bottom: schematic of the R‐ and S‐chains encoded in a single lentiviral vector, each separated by Venus or mScarlet. (G,H) Statistical quantification of NFAT‐GFP reporter activity (G) and IL‐2 production (H) in Jurkat T cells expressing stCAR HER2 or auxCAR HER2 co‐cultured with SK‐OV‐3 tumor cells in the presence of the indicated auxin concentration. n = 3 independent technical replicates (mean ± s.d.). (I) Statistical quantification of NFAT‐GFP reporter activity at the indicated time points following auxin removal in Jurkat T cells expressing auxCAR and co‐cultured with SK‐OV‐3 tumor cells. The auxCAR T cells were induced with 1 µM auxin.

Article Snippet: Recombinant human IL‐2 (Cat# RP01039, ABclonal) was added at a final concentration of 100 Unit/mL −1 to expand T cells.

Techniques: Flow Cytometry, Activation Assay, Expressing, Transduction, Cell Culture, Activity Assay, Concentration Assay, Plasmid Preparation

Journal: bioRxiv

Article Title: The Glucose Transporter GLUT3 Controls Regulatory T Cell Function

doi: 10.64898/2026.03.26.714439

Figure Lengend Snippet: (A and B) Relative body weight changes (A) and experimental autoimmune encephalomyelitis (EAE) clinical scores (B) in WT, GLUT1-deficient ( Slc2a1 fl/fl Cd4 Cre ) and GLUT3-deficient ( Slc2a3 fl/fl Cd4 Cre ) mice following immunization with MOG 35-55 peptide emulsified in CFA; means ± SEM of 6-11 mice per cohort. (C) RT-qPCR analysis of Slc2a1 (GLUT1) and Slc2a3 (GLUT3) expression in isolated splenic CD4 + T cells from WT, GLUT1-deficient and GLUT3-deficient mice used in EAE experiments; data are shown as means ± SEM of 4-11 mice. (D and E) Representative flow cytometric analysis of Foxp3 + Treg cells in the central nervous system (CNS) of WT, GLUT1- and GLUT3-deficient mice 20 days after MOG 35-55 immunization (D) , with quantification of Treg cell frequencies and absolute cell numbers (E) ; means ± SEM of 6-11 mice. (F and G) Frequencies of GM-CSF and IL-2-producing CD4 + T cells in the CNS of WT, GLUT1-and GLUT3-deficient mice 20 days after immunization with MOG 35-55 peptide and restimulation with PMA/ionomycin for 5 h (F) , with quantification of total cell numbers of GM-CSF and IL-2 producing CD4 + T cells in the CNS (G) ; means ± SEM of 6-11 mice. Statistical analyses shown in (A, B, C, E, G) were performed using two-way ANOVA. *, p<0.05; **, p<0.01; ***, p<0.001; ns, non-significant.

Article Snippet: FACS-sorted Treg cells were activated on Delta surface 96-well plates (Nunc) with 0.25 ng/mL anti-CD3 (clone 145-2C1) and 1 ng/mL anti-CD28 (clone 37.51; both Bio X Cell), 20 ng/mL recombinant human IL2 and 10 ng/mL murine IL7 for three days at a density of 2×10 6 cells/mL.

Techniques: Quantitative RT-PCR, Expressing, Isolation