Extended Data Fig. 4: AC484 enhances the activation of immune cell subsets and promotes immune infiltration of tumours.

a, Immunofluorescence microscopy of representative FFPE tumour sections from KPC untreated, anti-PD-1- or AC484-treated tumours, CD45 (green); CD8 (red); DAPI (blue). b, Quantification of CD45⁺ (left) and CD8⁺ (right) cells from KPC tumours from (a). c, Barplot showing the average normalized infiltration distance of CD45⁺ and CD8⁺ cells from the tumour border in immunofluorescence stains of KPC and B16 tumours. Infiltration distances are normalized as a fraction of the maximal distance found in each tumour. d, Schematic representing experimental design for single cell RNA sequencing on tumour-infiltrating lymphocytes from B16 or KPC tumours from control (n = 9), anti-PD-1- (n = 8) or AC484-treated (n = 10) tumours; figure created with BioRender.com. e, UMAP projection showing adequate batch correction of all cells with KPC recovered cells colored orange (top) and B16 colored blue (bottom). f, Heatmap showing top 15 differentially expressed genes in each cluster by one vs. rest test with two representative genes from each cluster shown on the y axis. g, Directional ratio of MDSCs versus monocytes by condition. h, In vitro CD86 expression (n = 6, left) and CXCL9 production (n = 4, right) by BMDM ± IFNγ. i, In vitro MHC I (left), MHC II (middle) and CD86 (right) expression on CD103⁺ BMDC ± IFNγ (n = 4). j, In vitro CXCL9 (left) and CXCL10 (right) production in CD103⁺ BMDC ± IFNγ (n = 4). k, In vitro IL12p70 and TNFα production by CD103⁺ BMDC ± IFNγ + anti-CD40 (αCD40, n = 4).

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