Fig. 5: CAR affinity and tumor antigen density regulate the degree of NFAT activation.

a Lentiviral vectors encoding C215-iGFP, UBS54-iGFP and Y6V-iGFP CARs. b Flow cytometric quantification of CAR-mediated NFAT activation, as indicated by NFAT-driven GFP expression in Jurkat CAR-T cells following stimulation for 24 h with target cells. CD69 was used as a marker for T-cell activation. Numbers indicate the percentages of live, single cells in the GFP- or CD69-positive gate. Neg, low, mid and high represent negative-, low-, medium-, and high-EpCAM expression, respectively. c Relative mean fluorescence intensity (MFI) of GFP and CD69 expression from b. Data were normalized to PMA/Ionomycin treatment and were presented as mean ± SD of quadruplicate samples.