Extended Data Fig. 6: APC-like tumour reprogramming induces an inflammatory profile in mouse cancer cells.
From: Intratumoural vaccination via checkpoint degradation-coupled antigen presentation
a, Confocal imaging of iVAC internalization in MDA-MB-231 cells using a FRET assay. Cy3 signal represents released OVA antigen; FRET signal indicates intact iVAC. Experiments were repeated independently two times. Scale bar: 10 μm. b, Following iVAC-induced antigenicity reprogramming, the leakage of delivered antigen from lysosomes for cross-presentation in tumours occurs via perforin-2 (Mpeg1). RNAi-mediated knockdown of perforin-2 can almost abrogate the antigen presentation by iVAC (n = 3 biological replicates). c–e, Heatmaps showing expression of genes related to (c) cell cycle, (d) IFN-γ and (e) STING pathways (n = 3 biological replicates). f, Gene set enrichment analysis (GSEA) for innate immune system genes, immune effector processes, and cytokine production in immune response genes in APC-like tumours derived from B16 cells compared to parental lines (HBSS, n = 3 biological replicates). Normalized enrichment score (NES) and false discovery rate q-values (FDR q) are provided. g,h, Quantitative PCR analysis of mRNA expression for antigen processing and presentation-related genes in APC-like tumours derived from (g) B16 and (h) MC38 cells (n = 3 biological replicates). In these experiments, the MC38 and B16 cells utilized were all murine PD-L1 knockout cells that expressed human PD-L1. Data are the mean ± s.d. Statistics: one-way ANOVA (b) and two-way ANOVA (g, h), each followed by Tukey’s multiple comparisons. NS: not significant.
