Extended Data Figure 2. H-2D^b mutant epitopes of d42m1-T3 tumours.

a, Missense mutations in d42m1-T3 were subjected to in silico analysis for the potential to form H-2D^b binding epitopes using three epitope prediction algorithms. The median predicted epitope binding affinity for each peptide was calculated and expressed as “median affinity value” where affinity value = 1/IC₅₀ x 100. Predicted epitopes are arrayed along the X-axis in alphabetical order based on their protein of origin b, Unfiltered median affinity values for the 4 predicted H-2D^b epitopes. c, Median affinity values of remaining 2 H-2D^b epitopes after filtering. d, Tetramer staining of CD8⁺ TILs from tumour bearing mice treated with αPD-1 using H-2D^b tetramers loaded with top 4 H-2D^b synthetic peptides. e, IFN-γ and TNF-α intracellular cytokine staining of CD8⁺ TILs from tumour bearing mice treated with αPD-1 immunotherapy following co-culture with naïve irradiated splenocytes pulsed with the top 4 H-2D^b synthetic peptides added at 1 μM final concentration. Data are presented as per cent of CD8⁺ TILs positive for IFN-γ, TNF-α, or both cytokines. Data are representative of two independent experiments.