Extended Data Fig. 2: IL-6/STAT3 signaling in hepatocytes remodels the tumor immune microenvironment.
From: Hepatocytes coordinate immune evasion in cancer via release of serum amyloid A proteins
a-c). Il6+/+ or Il6−/− mice were orthotopically (OT) injected with 5e5 PDA.69 cells. Twenty days later, tumors were harvested. n = 10 Il6+/+ mice and n = 9 Il6−/− mice were examined over 2 biological replicates. b) Tumors were formalin fixed, paraffin embedded, and stained for CD8 (brown), FOXP3 (purple), CK19 (yellow), and nuclei (blue, hematoxylin). Representative images are shown. Scale bars=100 μm. c) Quantification of CD8+ or FOXP3+ cells from (b). Mann Whitney tests were performed. d-g). C57BL/6 or AlbCre+/+Stat3fl/fl mice were orthotopically injected with 5e5 PDA.69 cells or received sham surgery. Twenty days later, livers and tumors were harvested for RNA sequencing. n = 6 mice per group. Data are representative of 1 biological replicate. d) Heatmap of differentially expressed genes (DEGs) in livers. e) Volcano plot showing DEGs in tumors of C57BL/6 or AlbCre+/+Stat3fl/fl mice. Genes enriched in C57/BL6 shown in blue. Genes enriched in the AlbCre+/+Stat3fl/fl group shown in red. Significance determined by log2FoldChange < −0.6 or > 0.6 and adjusted p value < 0.05. f) PCA plot showing clustering of tumors. g) Gene set enrichment analysis of DEGs. The ten most highly enriched gene sets in tumors of AlbCre+/+Stat3fl/fl mice are shown. h-k) C57BL/6 mice were OT injected with 5e5 T celllow (PDA.69) or T cellhigh (2838.c3) PDAC cells. Twenty days later, tumors were harvested. n = 5 mice per group. Data shown are representative of n = 1 biological replicate. i) Tumors were formalin fixed, paraffin embedded, and stained for CD8 (brown), FOXP3 (purple), CK19 (yellow), and nuclei (blue, hematoxylin). Representative images are shown. Scale bars=200µm. j) Quantification of CD8+ or FOXP3+ cells from (i). k) CD3+ T cell infiltration in tumors was analyzed by flow cytometry. l) C57BL/6 mice were treated with GFP.AAV or IL6.AAV as described in Fig. 2h, then injected with 5e5 T cellhigh (2838.c3) PDAC cells. IL-6 serum levels were measured at day 15 by cytokine bead array. Data are representative of 3 biological replicates. For j-l, Mann Whitney tests performed. For c, j, k, and l, data are presented as mean values +/- SD.
