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Figures
- Fig. 1.
Lentivirus specifically infects acinar cells in tree shrew pancreas. (A) Three animals were injected with EGFP-expressing lentivirus; green fluorescence was observed on day 4 around the injection site (red arrows). (B,C) Double labeling for GFP (green) and CK19 (red) (B) and GFP (green) and CPA1 (red) (C) in acinar cells. 4′,6-diamidino-2-phenylindole (DAPI; blue) was used to stain nuclei.
- Fig. 2.
Induction of pancreatic cancer in tree shrew. (A) Schematic illustration of lentiviral vectors. (B) shRNA-mediated knockdown efficiency in a tree shrew liver cell line. (C) Western blot analysis of KRASG12D, P53, P16 and P15 expression in tree shrew liver cells 4 days after lentiviral infection. (D) A tumor mass was observed at the head of the pancreas (red arrow). (E) Survival curve of the tree shrew pancreatic cancer model. The mortality of the KRAS-shTp53-shCdkn2a/b group was 100%, but there were no tumors or deaths recorded in animals of the KRAS-shTp53, KRAS-shTp53-shCdkn2a and KRAS-shTp53-shCdkn2b groups.
- Fig. 3.
Characterization of pancreatic cancer in tree shrew. (A,B) H&E staining of normal (A) and cancerous pancreatic tissue (B) from tree shrew. is, islet; black arrow, normal duct; red arrows, glandular structures; asterisks, fibrous stroma. (C,D) Active cellular proliferation, as shown by Ki67 immunoreactivity (red). (E-L) Immunohistochemical detection of human pancreatic cancer markers CK19 (E,F), Muc5 (G,H), MMP7 (I,J) and Hes1 (K,L) (red). (M,N) Mucin proteins were stained by Alcian Blue. Normal tissue, C,E,G,I,K,M; tumor tissue, D,F,H,J,L,N.
- Fig. 4.
Detection of ADM in early stages of pancreatic cancer development in tree shrew. (A-H) Tissue was harvested 7 days after injection of the lentivirus, and H&E staining was performed in the pTomo-empty (A), KRAS-shTp53 (B), KRAS-shTp53-shCdkn2a (C), KRAS-shTp53-shCdkn2b (D) and KRAS-shTp53-shCdkn2a/b (E-H) groups. H&E staining shows an increase in the number of ducts in tissue infected by lentivirus KRAS-shTp53-shCdkn2a/b; the areas in the blue line boxes are shown enlarged below. (I) Immunohistochemical analysis of CPA1 (red) and CK19 (green) expression in normal tissue and tissue infected by lentivirus KRAS-shTp53-shCdkn2a/b; arrows indicate double-positive cells.
- Fig. 5.
The Rb1 signaling pathway is involved in tree shrew pancreatic cancer. (A) Phosphorylation of Rb1 (Ser780) was detected by immunohistochemical staining in tree shrew pancreatic cancer tissue and normal tissue. (B) Phosphorylation of Rb1 (Ser780) was detected at the early stage of tree shrew pancreatic cancer; tissues were harvested 7 days after injection of pTomo-empty, KRAS-shTp53, KRAS-shTp53-shCdkn2a, KRAS-shTp53-shCdkn2b and KRAS-shTp53-shCdkn2a/b lentivirus.
- Fig. 6.
Comparisons of human, tree shrew and mouse pancreatic cancer models. (A) Homology comparison of ARF (encoded by Cdkn2a) among human, mouse and tree shrew. The homology of tree shrew to human was significantly higher than that of mouse to human. (B) Post-translational modification sites of P16Ink4a are outlined with red boxes, and the conserved sites in tree shrew, but not in mouse, are indicated by black arrows. ME, methylation; M1, mono-methylation; P, phosphorylation. (C) Pairwise comparisons of gene expression profiles of tree shrew, human and mouse PDAC. Dendrogram showing distinct clusters for the tree shrew, human and mouse samples. (D) Heat map illustrating the correlation between expression profiles of the three species. Hu, human; Mo, mouse; Ts, tree shrew. (E) Bar graph illustrating the average correlation among the three species; error bars represent s.d. ***P=1.25×10–25.
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