Fig. 3.

NUMB6-induced EMT phenotype increases DB-7 cancer cell migration and invasion in vitro. (A) Wound healing migration assay of DB-7 cells overexpressing GFP, NUMB4-GFP, or NUMB6-GFP. Cells as indicated were cultured until confluent and then a scratch wound was made using a 20ul pipette tip. Media were replaced and wounds were photographed at 20 hrs. (B) Graphic representation of migration differences between control GFP and NUMB4-GFP or NUMB6-GFP DB-7 cells. Error bars refer to SEM in pooled triplicate experiments. (C) Transwell migration assay of DB-7 cells overexpressing GFP, NUMB4-GFP, or NUMB6-GFP. Cells as indicated were seeded in transwell containing 8μm PET membrane and incubated for 20 h. The migrated cells were stained and scored. Error bars represent SEM. (D) Matrigel invasion assay of DB-7 cells overexpressing GFP, NUMB4-GFP, or NUMB6-GFP. Cells as indicated were plated in invasion chamber and incubated for 20 h. The transmigrated cells were stained and counted. The plotted data were averaged from three independent experiments and error bars represent SEM. Asterisks indicate significant differences between two groups.