Fig. 2.

Demonstration of antigen-specific responses against TOFU antigens. a–d Peptide stimulation and re-challenge of T-cells from Kluc TOFU mRNA primed animals (a), GL261 TOFU mRNA primed animals (b), NSC TOFU mRNA primed animals (c), and Ptch TOFU mRNA primed animals (d) with selected predicted neoantigens and TAA peptides (n = 3 per sample). IFN-γ release is detected using ELISA after 24 h. Antigen reactivity is considered as > 100 pg/ml IFN-γ production and at least a 2-fold increase over the response to the irrelevant peptide. Statistical analysis is done using 2-way ANOVA with multiple comparisons using Bonferroni correction (two-tailed); p < 0.05 is *, p < 0.01 is **, p < 0.001 is ***, and p < 0.0001 is ****. The experiments have been repeated at least 2 times to validate the immunogenicity