1l cysteine

The electric cell‐substrate impedance sensing (ECIS) system (Applied Biophysics, Troy, NY, USA) allows for electronic monitoring of cells on well‐arrays containing gold plated electrodes. As the cells cover the electrodes, impedance of current increases, which is measured as the output. A short pulse of a current 1000‐fold higher in magnitude causes cell death on the electrode, resulting in a 2‐D ‘wound’. The procedure was adapted from prior work 26. All experiments were performed on 96W1E+ ECIS arrays (Applied Biophysics). Prior to inoculation, the electrodes were cleaned with 10 mmol·L⁻¹l‐cysteine (pH 7.5, Sigma‐Aldrich) and rinsed twice with DI H₂O. HaCaTs were plated at 60 000 cells per well in 200 μL of full growth medium at 37 °C, 5% CO₂. After growing to confluence in 16–24 h, as shown by a plateau in impedance, the medium is switched to serum‐free DMEM with 1% P/S for 24 h at 37 °C, 5% CO₂. After serum starvation, each well received a ‘wound’ of 30 s at 5000 μA, 100 μL media was removed, and 100 μL treatments of LL‐37 and CSA‐13 were added at 2 ×  concentration. Migration response of impedance recovered over time was normalized to the control cells treated with serum‐free media. Data presented as mean ± SEM for N = 5 experiments.