Cell invasion assessment

Transwell chambers with Matrigel coating were utilized in order to perform invasion assays. Cells that underwent transfection were planted in the top chamber (1.0×10⁵ cells/chamber) and were incubated for 24 hours under conditions of 5% CO₂ and 37°C. FBS (20%) at the bottom of the chambers served as a chemoattractant A cotton swab was used to eliminate cells that did not exhibit invasion at the top surface after incubation, while those showing invasion at the bottom surface were fixed using 100% methanol and stained with 1% crystal violet. Microscope images were used to quantify the number of cells showing invasion characteristics. Six random fields were selected for quantification in each assay.