Figure 5.

PMCA4 knockdown inhibits cell migration without any effect on cell proliferation. MIA PaCa-2 cells seeded into Ibidi chambers were incubated with non-targeting control (siNT) or siRNA targeting PMCA4 mRNA (siPMCA4). After 24 h, the Ibidi chambers were removed to create a cell-free gap area at time 0. The anti-proliferative agent, 1 μg/ml mitomycin C (Mit C), was added after Ibidi chamber removal to prevent cell proliferation from influencing gap closure. (A) Representative images of the gaps at 0, 24, and 48 h. Gap areas (red outline) were processed and analyzed using ImageJ. (B) Data are presented as % gap closure with respect to time 0. (C) SRB proliferation assays were run in parallel to the migration assay to ensure cell proliferation was sufficiently inhibited by Mit C. Data are shown as mean ± SEM. (n = 4, 3–4 replicates per treatment condition). Comparisons were made between siNT control and siPMCA4 treated cells, with and without Mit C, at matching time points using two-way ANOVA with Tukey’s multiple comparison post-hoc test. * represents a statistically significant difference between % gap closure of siNT + Mit C and siPMCA4 + Mit C, at 48 h, where p < 0.05.