Extended Data Fig. 7: Inhibition of T cell surveillance by SAA depends on TLR2.
From: Hepatocytes coordinate immune evasion in cancer via release of serum amyloid A proteins
a) Study design for b. Saa+/+ or Saa−/− mice received sham surgery or orthotopic (OT) injection of 5e5 PDA.69 cells. n = 5 mice/group. Bulk liver tissue was collected in Trizol and RNA was extracted. b) Expression of known SAA receptors measured by qRT-PCR. Ordinary one-way ANOVA with Dunnett’s multiple comparisons test was performed. c-d) Twenty days after sham surgery or OT injection of 5e5 PDA.69 cells, livers were analyzed by flow cytometry. n = 5 mice per group. c) Percent of live cells expressing TLR2. d) tSNE plots were created from 5000 downsampled live cells and cell marker expression is displayed. For a-d, data are representative of n = 1 biological replicate. e-j) Tlr2+/+ or Tlr2−/− mice were OT injected with 5e5 PDA.69 cells. Twenty days later, tumors were harvested. Data are representative of 1 biological replicate for (f-g, j) and 2 biological replicates for (h-i). f) SAA serum levels at necropsy were analyzed by ELISA. n = 5 non-tumor bearing Tlr2+/+ mice, n = 9 Tlr2+/+ mice, and n = 10 Tlr2−/− mice. g) Livers were formalin fixed, paraffin embedded, and stained by RNA-ISH for Saa. h-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=100µm. n = 11 mice/group. j) T cell and dendritic cell (DC) infiltration into tumors was analyzed by flow cytometry. n = 8 Tlr2+/+ mice, n = 11 Tlr2−/− mice. For (c-j), significance was determined by Mann Whitney test. For b, c, f, h, and j, data are presented as mean values +/− SD.
