Panel A: Multiple line graphs show tumor size over time in mice bearing WT or TNFR1KO MC38 cells. The x-axis represents time in days, and the y-axis represents tumor size in square millimeters. The rightmost graph presents cumulative data with mean and standard error of the mean (SEM) and two-way ANOVA statistical comparisons. Panel B: Similar line graphs as in Panel A but with EO771 WT and TNFR1KO variants. The fractions in the top left corner of individual tumor graphs show the number of mice that experienced tumor rejection. Panel C: Line graphs show individual follow-up of tumor sizes in WT or Rag1 minus slash minus mice engrafted subcutaneously with the indicated WT or TNFR1KO MC38 variants. The right graph summarizes data with mean and SEM and provides two-way ANOVA statistical comparisons. Panel D: Line graphs show tumor size over time in mice inoculated with 1:1 or 1:5 mixtures of WT and TNFR1KO MC38 variants. The right graph presents cumulative data with mean and SEM and two-way ANOVA statistical comparisons. Panel E: Similar line graphs as in Panel D but with EO771 cells. Panel F: Line graphs show individual follow-up of right and left subcutaneous tumors in mice inoculated with MC38 WT or TNFR1KO variants along with a diagram. Panel G: Similar line graphs as in Panel F but with EO771 variants along with a diagram.
TNFR1KO tumor variants are controlled by CD8 + T lymphocytes, and small quantities of TNFR1 + co-injected variants locally rescue tumorigenicity of TNFR1KO tumor cells. (A) Selective T cell depletion experiments in mice bearing WT or TNFR1KO MC38 cells treated with control antibody or depleted of CD4+ or CD8β+ lymphocytes, with the lower graph presenting cumulative data (mean ± SEM) and two-way ANOVA statistical comparisons (n = 6). (B) Results as in A but with EO771 WT and TNFR1KO variants. Fractions in the top left corner of individual tumor graphs show the number of mice that experienced tumor rejection (n = 6). (C) Individual follow-up of tumor sizes in WT or Rag1−/− mice engrafted subcutaneously with the indicated WT or TNFR1KO MC38 variants. The graph on the right summarizes data (mean + SEM) and provides two-way ANOVA statistical comparisons (n = 5–12). (D and E) Experiments as in Fig. 1 but inoculating either 1:1 or 1:5 mixtures of the WT and TNFR1KO variants, using MC38 (D) or EO771 (E) cells, respectively. Cumulative data (mean ± SEM) and two-way ANOVA statistical comparisons are presented in the figures (n = 6). (F and G) Experiments testing subcutaneous contralateral inoculation of tumor cells, either WT (represented in black) or TNFR1KO (represented in light brown). Individual follow-up of right and left subcutaneous tumors is shown. F refers to MC38 variants, while G corresponds to EO771 variants. The shown experiments are representative of two replicates, with the exception of F and G, which represent a single experiment. P < 0.05 (*), P < 0.01 (**), P < 0.001 (***), and P < 0.0001 (****).