Phospho-LAT (Tyr161) Antibody - #AF8200

Fig. 3 CD3ε-based CAR-T cells produce low levels of inflammatory cytokines. a–c T cells engineered with CARs containing varied ICDs were cocultured with the corresponding malignant cells overexpressing HER2 (a), mesothelin (b) or CD19 (c) (E:T = 1:1) for 24 h. The levels of indicated cytokines produced by CAR-T cells were measured and plotted. d–f CAR-T cells were cocultured with indicated malignant cells that expressed the corresponding antigens (E:T = 1:1) for 24 h, and the percentages of cell lysis were calculated and plotted. “19” in the CAR names refer to an scFv against CD19. g,h Normalized enrichment scores of significantly changed gene sets in H28E versus other groups of CAR-T cells as determined in Gene Ontology (g) analyses (n = 3 replicates per group). The enrichment of gene sets related to cytokine or chemokine activity between H28Z and other groups of CAR-T cells was also determined by GSEA (h; n = 3 replicates per group). i HER2-targeted CAR-T cells were stimulated by incubation with HER2-overexpressing PC-9 cells for indicated times. Cells were then harvested for Western blot analysis. The intensities of the protein bands were quantified by the ImageJ software, and the normalized band densities were indicated. Data are representative images and expressed as the means ± SD of three independent experiments. **P