Growth of Snail-expressed Panc-1 cells in three different conditions.

(A) Monolayer culture. Mock-Panc-1 (open column) and Snail-Panc-1 (closed column) were seeded at a density of 1.0×10⁴ cells per well of 24-well plates in 10% FBS-containing medium and incubated for 7 days. After the incubation, the number of cells was measured with a cell counter. (B–D) Soft agar culture. Mock-Panc-1 and Snail-Panc-1 cells were cultured in soft agar medium for 14 days. After the incubation, the number of total colonies (B) and total colony area (C) were analyzed by Image J. The soft agar cultures were photographed and each typical image is shown in (D). Scale bar, 500 µm. Other experimental conditions were the same as described in Figure 4. (E) Collagen culture. Mock-Panc-1 and Snail-Panc-1 cells were cultured in 3D collagen gel for 7 days as described in Figure 5. After the incubation, the relative number of cells was measured using Dojindo cell counting kit 8.