PancTu-I shTR2 cells show reduced cell growth in the presence of HSC in vitro. PancTu-I shCtrl or PancTu-I shTR2 cells were indirectly cocultured in the presence of HSC or HMF. After 6 days, (A) the vital cell number, (B) the proportion of PI+ cells, and (C) Ki67+ cells were determined. Vital cell numbers were obtained by counting living cells in a Neubauer counting chamber. The fraction of cells which underwent cell death was quantified by PI staining. The percentage of proliferating cells was determined by immunocytochemical Ki67 staining. (D) Representative images of Ki67 staining in PancTu-1 cells after different coculture conditions at 200-fold magnification. For extended coculture, HSC cocultured PancTu-I shCtrl and PancTu-I shTR2 cells were detached and re-seeded to extend the coculture either in the presence of HSC (HSC→HSC) or HMF (HSC→HMF). After another 6 days of coculture, (E) vital cell count and (F) the proportion of Ki67+ cells were assessed. The percentage of proliferating cells was determined by immunofluorescence Ki67-Alexa-488 staining. Data are expressed as median fluorescence intensity (MFI). Data represent the mean ± SEM of 3–5 independent experiments; * = p < 0.05.