Figure 4. The 1^st‐generation CAR forms microclusters and immunological synapses in LAT‐deficient Jurkat T cells.

1. Schematics of 3^rd and 1^st generation of CAR
2. WT or LAT‐deficient (Jcam2.5) cells expressing the 3^rd‐ or 1^st‐generation CAR were stimulated on supported lipid bilayers coated with CD19 and ICAM‐1. TIRF microscopy revealed clustering of CAR and Ax647‐labeled streptavidin–biotin‐CD19. Scale bar, 2 μm.
3. Quantification of clustering level of Ax647‐labeled streptavidin–biotin‐CD19. Shown are means ± SD. Clustering is quantified as normalized variance. N = 50 cells for CAR 3^rd WT and 1^st WT, 63 cells for CAR 3^rd lat‐, and 51 cells for CAR 1^st lat‐. **: 0.0005 < P < 0.005. Statistical test: unpaired two‐tailed t‐test.