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Fig. S1. (a) Propidium iodide (PI) fluorescence intensity of green fluorescent protein (GFP)-SUIT-2 cells. (b) The GFP fluorescence intensity was correlated with the PI fluorescence intensity (Spearman’s rank-correlation coefficient: 0.998, < 0.0001). (c) GFP-SUIT-2 cells expressed similar levels of fluorescence intensity to monocultured cells, regardless of the number of co-cultured pancreatic stellate cells (PSCs). (c,d) Regression analysis confirmed that the fluorescence intensity of GFP-SUIT-2 cells was correlated with the number of cells, despite the coexistence of myofibroblasts and their morphological alterations (Spearman’s rank-correlation coefficient: 0.993, < 0.0001).

Fig. S2. (a) Representative microphotographs of green fluorescent protein (GFP)-SUIT-2 cells in monoculture (left), indirect co-culture with MRC5 fibroblasts (center), and direct co-culture with MRC5 fibroblasts (right). Monocultured GFP-SUIT-2 cells were almost round in shape, whereas co-cultured cells exhibit a fibroblastoid morphology. (b) GFP-expressing cancer cells and GFP-negative MRC5 fibroblasts were isolated using a cell sorter. (c) The Notch-1 levels were only elevated in directly co-cultured MRC5 fibroblasts. (d,e) The hairy and enhancer-of-split homolog-1 (Hes-1) and Jagged-1 mRNA levels in directly co-cultured cells were dramatically elevated compared with those in indirectly co-cultured and monocultured cells.

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