Fig. 2

Pancreatic cancer cells promote SC dedifferentiation. a Schematic diagram of the coculture system of PDAC cells and SCs. b Venn diagram of upregulated pathways in RSC96 (+PANC-1) cells and RSC96 (+BxPC-3) cells compared with RSC96 cells (n = 3). c, e Transwell assays were used to evaluate the effects of PANC-1 and BxPC-3 cells on the migration of sNF96.2 cells (n = 6). Scale bar: 100 μm. d Microscopy images showing the morphological changes in sNF96.2 cells induced by PANC-1 and BxPC-3 cells. Scale bars: 50 μm (upper panels) and 20 μm (lower panels). f–j Western blotting was performed to detect the protein expression levels of dedifferentiation markers (p75NTR, c-Jun, SOX2, N-cadherin, and GDNF) in sNF96.2 cells after coculture with PANC-1 or BxPC-3 cells (n = 3). k, m Transwell assays were used to assess the effects of PANC-1 and BxPC-3 cells on the migration of RSC96 cells (n = 6). Scale bar: 100 μm. l Microscopy images showing the morphological changes in RSC96 cells induced by PANC-1 and BxPC-3 cells. Scale bars: 50 μm (upper panels) and 20 μm (lower panels). n–r Western blotting was conducted to determine the protein expression levels of dedifferentiation markers (p75NTR, c-Jun, SOX2, N-cadherin, and GDNF) in RSC96 cells after coculture with PANC-1 or BxPC-3 cells (n = 3). Relative expression levels were quantified as p75NTR/GAPDH, c-Jun/GAPDH and GDNF/GAPDH; since the molecular weight of SOX2 (~35 kDa) is close to that of GAPDH (~36 kDa), SOX2/β-actin was used for relative expression quantification. All the results are presented as the means ± SDs. Each data point in the bar graphs represents an individual sample. Statistical significance was determined by one-way analysis of variance followed by Tukey’s HSD post hoc test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001