Ecis system software

For impedance measurements, an ECIS instrument model Z (Applied Biophysics, Troy, NY, USA) was used. To start the trials, electrodes (pre-treated with aqueous 10 mM cysteine solution as recommended by the manufacturer) were filled with 400 µL of DMEM. HeLa cells were seeded at 1 × 10⁴ cells/well on electrode plates, and after a 24 h incubation, cells were allowed to grow in 2% FBS-containing medium containing 5 ng/mL TGF-β1. The entire electrode array was placed in a plate holder connected to the ECIS system and cell proliferation analysis was performed using the ECIS system software (Applied Biophysics). A growth curve was plotted based on cell proliferation percentage and time.

ECIS^® (Electric Cell-substrate Impedance Sensing) is a real-time, label-free, impedance-based method of assessing the activities of cells grown in tissue culture. When cells are added to the ECIS Arrays and attached to the electrodes, they act as insulators and increase the impedance. As cells grow and cover the electrodes, the current is impeded in a manner related to the number of cells covering the electrode, the morphology of the cells, and the nature of the cell attachment. When cells are stimulated to change their function, the accompanying changes in cell morphology alter the impedance. Data are generated as impedance versus time.
Electrode plates (eight wells, Applied Biophysics, Troy, NY, USA) were induced with an electrode-stabilizing solution provided by the manufacturer 10 min prior to seeding with HaCaT cells overnight. Cells were then treated with different concentrations of DK223. After 24 h, the electrode plates were placed in the ECIS apparatus in the incubator for migration analysis using the ECIS system software, according to the manufacturer’s instructions (Applied Biophysics, Troy, NY, USA).