Figure 8.

Effects of intracellular Ca²⁺ and SOCE activation on spreading of MIN6 cells attached to E-cad/Fc. (A) The protocol to determine whether intracellular Ca²⁺ signaling is important for cell-cell adhesion. (B) MIN6 cells were seeded on glass slides coated with 5 μg/ml E-cad/Fc for 6 h, and then treated with or without fluoxetine for 3 h. After one-hour incubation with fluoxetine, cells were treated with or without ionomycin (5 μM) for 2 h. Cells were rinsed with PBS to remove unattached cells, and then visualized by EVOS FL Auto. At least 100 cells were analyzed for each cell population. Scale bar, 50 μm (C) MIN6 cells were seeded on glass slides coated with 5 μg/ml E-cad/Fc for 4 or,7 h, and then treated with or without SKF-96365 (25 μM or 50 μM) for 2 h. Cells were rinsed with PBS to remove unattached cells, and then visualized by EVOS FL Auto. At least 100 cells were analyzed for each cell population. Scale bar, 50 μm (D) Quantitative analysis of the percentages of spreading cells. Each value represents mean ± SEM from at least three independent experiments. ***p < 0.001 (E) Quantitative analysis of the percentages of spreading cells. Each value represents mean ± SEM from at least three independent experiments. ***p < 0.001.