Extended Data Fig. 1: Liver- and Lung-avid cell lines exhibit distinct features.
From: PCSK9 drives sterol-dependent metastatic organ choice in pancreatic cancer
a, b. Petal plots depicting metastatic potential (length) and penetrance (width) for C1-Liver lines (a) and C2-Lung lines (b), extracted from the MetMap 500 analysis. Br, Brain; Bo, Bone; Ki, Kidney; Li, Liver; Lu, Lung. c. Representative H&E images showing tumor growth (red outline) in the liver (top) and lung (bottom) 3 weeks post intra-cardiac injection of the indicated PDAC cell lines. Scale bars: 500 µm. d. Quantification of liver/lung colonization ratio of C1-Liver and C2-lung lines for the experiment in c. (n = 6 mice per/cluster). Statistical significance was calculated using a two-tailed t-test. e. Flow cytometry-based measurement of relative eGFP or mCherry enrichment in the liver or lungs. mCherry positive cells were more abundant in the liver while eGFP positive cells were more abundant in the lungs (related to Fig. 1l–n). Statistical significance was calculated using a two-tailed t-test. Error bars depict standard deviation. (n = 3 tissues per condition). f. Gene set enrichment analysis (GSEA) plots showing relative enrichment of the PDAC classical gene signature derived from Adams et al. and Moffit et al. in cluster 1 (C1-Liver) and cluster 2 (C2-Lung) cell lines. Statistical significance was calculated using a permutation test with FDR adjustment. g. Scatter plot showing correlation between liver/lung metastatic potential ratio (see Supplementary Table 2) and classical score from Adams et al. and Moffit et al. Each dot represents a PDAC cell line (n = 25). Linear regression followed by Pearson correlation. Error margins (grey) depict standard error of the mean. h. Representative immunofluorescence images of GATA6 (green), Pan-Cytokeratin (CK19) (red), DAPI (blue) in liver (top) and lung (bottom) patient PDAC metastases. Note the nuclear localization of GATA6 in lung metastases. Scale bars: 20 µm. i. Quantification of the percentage nuclear GATA6 in liver (56 fields from n = 3 samples) and lung (45 fields from n = 3 samples) metastases sections. Statistical significance was calculated using a two tail T-test. j. Representative H&E images of human liver and lung PDAC metastases. Scale bar: 20 µm. k. Bar plot depicting quantification of the differentiation status of PDAC liver and lung metastases shown in j, (Liver metastases n = 45; Lung metastases n = 10). Statistical significance was calculated using a two-tailed t-test. l. Representative immunofluorescence images of ZEB1 (green), Pan-Cytokeratin (CK19) (red), DAPI (blue) in liver and lung patient PDAC metastatic lesions. Note the nuclear localization of ZEB1 in the liver metastases. Scale bars: 20 µm. m. Quantification of the percentage nuclear ZEB1 in liver (55 fields from n = 5 samples) and lung (40 fields from n = 5 samples) metastases sections. Statistical significance was calculated using a two tail T-test. For flow cytometry source data, see Supplementary Fig. 2.
