Supplementary Figure 1: Morphological characterisation of metastatic PDAC lesions in human and mouse.
From: Macrophage-secreted granulin supports pancreatic cancer metastasis by inducing liver fibrosis
(a) Lower magnification of representative micrographs shown in Fig. 1a. Identification of pan-cytokeratin (CK)+ metastatic pancreatic cancer cells, hematopoietic cells (CD45+), macrophages (CD68+) and myofibroblasts (αSMA+) as predominant cell types at the hepatic metastatic microenvironment of pancreatic cancer by immunohistochemical analysis of human biopsies (data are from 5 PDAC patients and 5 healthy subjects; five fields assessed per sample). HL = healthy liver, LM = liver metastasis. (b) Identification of cytokeratin (CK) 19+ metastatic pancreatic cancer cells, tumour associated macrophages (CD68+) and myofibroblasts (PDGFRα+) as predominant cell types at the hepatic metastatic microenvironment of pancreatic cancer by immunofluorescence analysis of human biopsies. Representative micrographs are shown and quantification of data (n = 5 PDAC, n = 5 healthy subjects; five fields assessed per sample; mean ± s.e.m.; two-tailed unpaired t-test). HL = healthy liver, LM = liver metastasis. (c) Representative Masson’s trichrome staining and quantification of area occupied by fibrotic stroma in human biopsies (n = 5 PDAC patients, n = 5 healthy subjects; five fields assessed per sample; mean ± s.e.m.; two-tailed unpaired t-test). HL = healthy liver, LM = liver metastasis. (d) Experimental metastasis model by intrasplenic implantation of 1 × 106 KrasG12D; Trp53R172H; Pdx1-Cre (KPC) mice derived pancreatic cancer cells expressing a luciferase/zsGreen lentiviral reporter plasmid (KPCluc/zsGreen). Liver tissues were isolated at day 5 and day 12 post implantation. (Upper panel) Liver tissue sectioned stained by Hematoxylin and Eosin (HE) showing initial micrometastatic lesions of disseminated cancer cells at day 5 post implantation followed by the generation of an excessive stromal microenvironment surrounding disseminated cancer cells at day 12 post implantation. (Lower panel) Representative immunofluorescence staining of disseminated KPCluc/zsGreen (zsGreen) cells. Data are from 6 mice per time point; four fields assessed per sample; data combine two independent experiments. Scale bars = a, 200 μm; b,c,d, 100 μm.
