Extended Data Fig. 2: hHER2 CAR-T cell distribution.
From: ST3GAL1 and βII-spectrin pathways control CAR T cell migration to target tumors
A. Deep 3D imaging (1 mm thick) of CAR-T cell accumulation in the cleared mouse lung after 72 hr after i.v. injection (red; CD11c or MHCII, green; CAR-T cell). Graph shows colocalization of red signal versus GFP (CAR-T cells). Pearson coefficient was generated as Green (CAR-T cell) /Red (anti-CD11c or anti-MHCII). Data are presented as mean ± SEM; n = 3 mice/group. B. Flow cytometric analysis of OT-I T cells in the lung, blood, and tumor after i.v. injection (4, 24, 48, and 72 hr; mean ± SEM, n = 3 mice per group). C. PCA of differentially expressed genes in CAR-T cells (T72 vs. T0). D. Gating strategy for detection of hHER2-CAR-T cells from organs (lungs) E. Expression of Ki67+ in CAR-T cells isolated from the lung after i.v. injection. F. Relative expression of IFN-γ and TNF in CAR-T cells isolated from lung at indicated times after i.v. injection. Data are presented as mean ± SEM; n = 3 mice/group. G. Representative flow cytometry results show cell surface PD-1 and Tim-3 expression levels in OT-I naïve T cells, freshly activated in vitro (0 hr), or isolated from the lung after 72 hr post-injection. H. hHER2-CAR-T cells isolated from the lung or dLN after 72 hr i.v. injection were cocultured with B16-hHER2 for 24 hr. Dead B16 cells were stained with 7-AAD. The data reflect 3 independent experiments (the mean ± SEM, n = 3). The data were analyzed by two-tailed Student’s t test (*P = 0.002). I. Representative flow cytometry results showing cell surface PD-1 and Tim-3 expression levels in naïve and CAR-T cells isolated from the lung at the indicated times post-injection.
