Figure 3.

Neurite growth in Transwell co‐culture of PC12 cells with PDAC cell lines. PC12 co‐cultured on their own was used as controls. NGF (50 ng·mL⁻¹) treatment was used as a positive control. Representative images of PC12 neurite growth in co‐culture with PDAC cell lines (A). Cells extending neurites longer than the maximum diameter of the cell body were counted and represented as a percentage of total number of cells (B). PC12 survival in transwell co‐cultures using Annexin V/DAPI flow cytometry assay. SF (serum‐free media) conditions were used as positive control for apoptosis, whereas CM (complete media) and NGF (50 ng·mL⁻¹) were negative controls. Following 24 h co‐culture in serum‐free‐media, floating and attached PC12 cells were harvested, stained with Annexin and DAPI, and analyzed as detailed in Materials and methods section (C). The percentage of live cells (negative for both Annexin V and DAPI) in each condition was expressed relative to live cells in complete media (D). Western blot showing lower expression of Caspase 3 (E); scale bar = 50 μm; 3B/D, ANOVA, ***P < 0.001; error bars indicate SEM; n = 3.