Extended Data Fig. 8. Signal 1 enhancement improved the accumulation, persistency, and effector function of iCAR-TCTL.
a. The gRNA sequences of intact DGKα (exon 7) and DGKζ (exon 2) genes (upper and middle panels). Sequences of mutated DGKα and DGKζ genes in both alleles in DGK-dKO-CAR-iPSCs (lower panels). b. Western blotting for DGKα and DGKζ in iCAR-TCTL and DGK-dKO-iCAR-TCTL by a capillary electrophoresis-based Protein Simple Wes System. Data were processed using the Compass software (Protein Simple, San Jose, CA, USA). c. Ratio of CD5CD8β DP cells to live differentiated cells derived from wild-type iCAR, DGKα-KO iCAR, DGKζ-KO iCAR, DGK-dKO iCAR, and DGKα-/-DGKζf/f CAR-iPSCs. c. Surface antigen profiles of iCAR-TCTL, DGK-dKO-iCAR-TCTL, and pCAR-TCTL. iCAR-TCTL, DGK-dKO-iCAR-TCTL, and pCAR-TCTL were evaluated using surface antigen expression related to T/NK linage, naïve/memory, and exhaustion phenotype. d. Detection of tumor-infiltrating T cells by FCM in JHH7-bearing xenograft mice treated with iCAR-TCTL or DGK-dKO-iCAR-TCTL. e. Tumor volume of inoculated JHH7 at indicated time points in individual mice treated with indicated test cells. Mean tumor size ± SEM (n = 8 of each group). Two-way ANOVA. n.s., not significant; SEM, standard error of the mean.