Fig. 4. CTL-mediated immune selection enriches TCTP⁺ immune-refractory tumor cells.

a Schematic of the therapy regimen in NOD/SCID mice implanted with A375 P0 or P3 cells. b TPT1 mRNA and protein levels at various stages of immune-resistance A375 cells were determined by qRT-PCR and Western blot. c TCTP⁺ tumor cells were analyzed by flow cytometry. d The protein levels of CXCL10 and MCL-1 were analyzed. e–g P3 cells were transfected with the indicated siRNAs. e The protein levels of TCTP, CXCL10, and MCL-1 were determined by Western blot. f Transwell-based T cell chemotaxis assays were performed using siGFP- or siTPT1-treated A375 P3 cell-derived CM. g CFSE⁺ apoptotic tumor cells (active-caspase-3⁺) was determined by flow cytometry. h Schematic of the therapy regimen in NOD/SCID mice implanted with A375 P3 cells. i–m P3 tumor-bearing mice administered siGFP-or siTPT1-CNPs with or without NY-ESO1-specific T cell adoptive transfer. i Flow cytometry profiles of CFSE⁺ adoptively transferred NY-ESO1-specific T cells. j Active-caspase-3⁺ apoptotic cells in the tumors. k The frequency of apoptotic cells in the tumor relative to the NY-ESO1-specific T cells migrated to the tumor. l Tumor growth and m survival of mice inoculated with A375 P3 cells treated with the indicated reagents. All in vitro experiments were performed in triplicate. For the in vivo experiments, 10 mice from each group were used, and randomly selected 5 samples were analyzed i–k. The numbers below the blot images indicate fold change b, d, and e. The p values by one-way ANOVA b, c, j, and two-tailed t test f, i, k, and two-way ANOVA g, l, and the log-rank (Mantel–Cox) test m are indicated. In the box plots, the top and bottom edges of boxes indicate the first and third quartiles; the center lines indicate the medians; and the ends of whiskers indicate the maximum and minimum, respectively. The data represent the mean ± SD. Source data are provided as a Source data file.