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Figure S1. The immunofluorescent staining for the expression of self-renewal regulators in pancreatic cancer cells (100×). The human pancreatic cancer cell lines and their sphere-forming cells were stained with DNA dye (Hoechst33342) or specific antibodies for the expression of OCT4 (green) and Nanog (red). Pc, parental cells; Sc, sphere cells.

Figure S2. The immunofluorescent staining for the expression of Notch2 in Bxpc-3 and Panc-1 spheres (200×). (A–C) Bxpc-3 spheres. (D–F) Panc-1 spheres. (A,D) Tumourspheres observed under an optical microscope. (B,E) The tumourspheres were stained with DNA dye (Hoechst33342). (C,F) The green fluorescence indicated the Notch2.

Figure S3. The immunofluorescent staining for the expression of stemness transcription factors in Panc-1 and Bxpc-3 cells (×200). The pancreatic cancer Panc-1 and Bxpc-3 cells were stained with DNA dye (Hoechst33342) and specific antibodies (red) for the expression of OCT4, Nanog and PDX1. The positive expressions of OCT4, Nanog and PDX1 in Notch2+ Panc-1 and Bxpc-3 cells; the positive expression of Nanog in Notch2 Panc-1 and Bxpc-3 cells, but the expressions of OCT4 and PDX1 were negative in these cells.

Figure S4. The immunohistochemical staining of Notch2 in pancreatic tissues. Immunohistochemical staining was used to detect the expression of Notch2 in normal pancreatic tissues (A–D) and pancreatic carcinoma tissues (E–H). The membranes of Notch2+ cells were stained dark brown. (A) A small proportion of cells were Notch2 positive in normal pancrease tissues (100×). (B) The Notch2+ cells were concentrated at the pancreatic terminal duct (200×). (C) The centroacinar cell (red arrow) at the point of pancreatic terminal duct (yellow arrow) was Notch2 positive (400×). (D) Pancreatic acinar cells and centroacinar cell (red arrow) were shown by H&E stain (400×). (E) Some sporadic Notch2+ cells were presented in the poorly differentiated pancreatic carcinoma tissues without glandular cavity (200×). (F) A partial enlarged view of diagram ‘E’. The positive expressions of Notch2 were distributed in the membranes of some pancreatic cancer cells (red arrow) (400×). (G) The pancreatic cancer cells (red arrow) located in the infiltrated nervus (yellow arrows pointed out the edge of a nervus) were illustrated (100×). (H) A partial enlarged view of diagram ‘G’. The Notch2+ cells were aggregated in the invasive front of pancreatic cancer (200×).

Figure S5. The viability of Notch2+Bxpc-3 cells after the treatment with CE.

Table S1. In vivo tumourigenicity of Notch2+/Notch2 Bxpc-3 pancreatic cancer cells.

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