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: medRxiv

Article Title: Targeting PTP1B and DUSP4 phosphatases to Boost Tregs: A Novel Therapy for Polyendocrine Metabolic Ovarian Syndrome (PMOS) Immune Dysfunction

doi: 10.64898/2026.05.30.26354518

Figure Lengend Snippet: Plasma level IL-2 in control (n=37) and PMOS (n=37) subjects and altered gene expression of JAKs in the total leukocyte RNA of 102 control and 102 PMOS subjects. A: Dot plot graphical representation of IL-2 level in the control and PMOS subjects. B-D: Dot and violin plot graphical representation of real-time gene expression level in JAK1, JAK2 and JAK3 . Each dot represents an individual subject. Each reaction was performed in triplicates and the ΔCt method was used to calculate the gene expression difference between control and PMOS subjects. 2 −ΔCt values of each subject are plotted on the Y-axis of the graph. Unpaired student’s T-tests were performed using GraphPad Prism 8 and p-value< 0.05 were set to be significant. Graph E and F: Dot and bar graph representation of relative gene expression level of JAK1, and JAK2 among the control and PMOS phenotype A, B, C, and D. The control group represented black dots, phenotype A in blue dots, phenotype B in orange dots, phenotype C in violet dots and phenotype D in yellow dots. One-way ANOVA was performed using GraphPad Prism 8 and p-value< 0.05 were set to be statistically significant.

Article Snippet: After 24 hours of siRNA treatment, the cells were stimulated with 300IU of recombinant IL-2 (rIL-2) (#130-097-743, Miltenyi Biotech, Germany).

Techniques: Clinical Proteomics, Control, Gene Expression

Journal: medRxiv

Article Title: Targeting PTP1B and DUSP4 phosphatases to Boost Tregs: A Novel Therapy for Polyendocrine Metabolic Ovarian Syndrome (PMOS) Immune Dysfunction

doi: 10.64898/2026.05.30.26354518

Figure Lengend Snippet: FACS analysis showed an increase of CD4 +ve CD25 +ve CD127 low/- Treg population after PTP1B and DUSP4 (MKP-2) siRNA-based knockdown in PBMCs of PMOS (n=12) subjects. A: bar graph representation showing the gene expression of PTP1B and DUSP4 across the PTP1B, DUSP4, and both silenced PBMCs. B: Dot plot and histogram representation of gating strategy with P1 shows lymphocyte population, P2 shows CD4-FITC +ve population, P3 shows CD25-PE Cy7 +ve population and P4 shows CD127 low/- Treg population. Gating is based on histogram of unstained and stained population (FITC, PE Cy7 and Alexa Fluor 647 respectively). C: Dot plot representation of CD4-FITC +ve CD25-PECy7 +ve CD127-Alexa Fluor 647 low/- (P4) Treg population of all treatment conditions including; untreated, IL-2 treated, control siRNA silenced, control siRNA silenced with IL-2, PTP1B siRNA silenced without IL-2 and with IL-2, DUSP4 siRNA silenced without IL-2 and with IL-2, and PTP1B+DUSP4 siRNA silenced without IL-2 and with IL-2. D: Dot and truncated violin plot representation of the percentage of CD4 +ve CD25 +ve CD127 low/- Treg population in lymphocyte events across all the treatments. Each dot represents an individual subject. One-way ANOVA was performed using GraphPad Prism 8 and p-value< 0.05 were considered as statistically significant.

Article Snippet: After 24 hours of siRNA treatment, the cells were stimulated with 300IU of recombinant IL-2 (rIL-2) (#130-097-743, Miltenyi Biotech, Germany).

Techniques: Knockdown, Gene Expression, Staining, Control