Journal: Nature Immunology

Article Title: Fine-tuning BACH2 dosage balances stemness and effector function to enhance antitumor T cell therapy

doi: 10.1038/s41590-025-02389-z

Figure Lengend Snippet: a , Experimental schema. OT-I splenocytes were activated for 24 h with anti-CD3 and anti-CD28 before retroviral transduction. Transduced cells were maintained in IL-2-supplemented media on plates coated with (chronic stimulation) or without (acute stimulation) anti-CD3. b , Representative flow cytometry illustrating PD-1 and TIM-3 expression on EV-transduced OT-I cells following acute or chronic stimulation. c , d , Percentage of PD-1 + TIM-3 + chronically stimulated ( c ) or IFNγ + TNF + acutely stimulated following 4 h of anti-CD3 restimulation in the presence of brefeldin A and monensin ( d ) from OT-I T cells transduced with the indicated vectors ( n = 5 for all groups) at day 4 and representative flow cytometry plots. e , FSC-A of acutely stimulated OT-I T cells transduced with the indicated vectors ( n = 5 for all groups) at day 4 and representative flow cytometry histograms. f , Heat map showing differentially expressed genes (DEGs, q < 0.05, log 2 (fold change or FC) > 1) between chronically stimulated transduced OT-I T cells ( n = 4 for all groups). Color indicates row z score. Black bars indicate genes bound by BACH2 based on a prior chromatin immunoprecipitation sequencing (ChIP–seq) analysis . g , Alignment showing representative mRNA expression at indicated loci within EV, BACH2 OE , BACH2 DE-10% and BACH2 DE-5% of Cxcr6 , Ccr2 and Havcr2 ; ChIP–seq analysis of BACH2 binding and ATAC-seq analysis of chromatin accessibility. Black arrowheads represent AP-1 binding motifs (TGA(G/C)TCA) colocalizing with BACH2 binding peaks. Data are representative of three independent experiments ( a – e ) with three to five samples per experimental group in each experiment. One-way ANOVA with Dunnett’s multiple-comparison correction ( c – e ). Dots represent independent replicates ( c – e ); horizontal lines and error bars indicate the mean ± s.e.m ( c – e ).

Article Snippet: Platinum-E retroviral ecotropic packaging cells (Plat-E) were purchased from Cell Biolabs.

Techniques: Retroviral, Transduction, Flow Cytometry, Expressing, ChIP-sequencing, Binding Assay, Comparison