Figure 2.

Functional role of hERG1 channels in PDAC cells. (A) Effect of E4031 on PDAC cell proliferation. PANC-1, MIAPaCa-2 and BxPC-3 cells were seeded at 10 000 cells per well and treated at time zero with E4031 (40 μM, final concentration). The number of viable cells were estimated through the trypan blue exclusion method at different times of incubation. (B) Effect of E4031 on PDAC cell growth in soft agar. When PDAC cell lines were cultured in soft agar, clonal growth was inhibited significantly by hERG1-specific inhibitor E4031. Colonies were morphologically similar and reached the same size. For PANC-1 and MIAPaCa-2 cell lines 3000 cells per plate were seeded, whereas for BxPC-3 cell line 5000 cells per plate were seeded. (C) Effect of E4031 on migration of PDAC cells. The cell trajectories are normalised using a common starting point. Each track represents the movement of one cell during the 5 h period. The radii of the circles represent the mean distances covered within ∼5 h. The method is fully described in Supplementary Information (Materials and Methods of in vitro experiments). *P<0.05; **P<0.02; ***P<0.01 (Student's t-test). (D) E4031 effects on primary PDAC cells. a, b: representative microphotographs of a primary PDAC cell culture. Left panels: control cells; right panels: E4031-treated cells. Magnification: 200x (a) and 400x (b). In panel c, a histogram showing cell viability (evaluated by WST-1 assay, as described in Supplementary Information) in control (black bar) and E4031-treated cells (grey bar) is reported.