HaCaT cells (immortalized human keratinocyte line) (CLS cell lines service, Germany, Cat nº 300493) were cultured in Dulbecco’s Modified Eagle Medium (Gibco, Gaithersburg, MD, USA), supplemented with 10% fetal bovine serum (Gibco) and 50 µg/mL of gentamicin (Gibco) at 37 °C and 5% CO2. HaCaT monolayers on a 24-well plate were scratched and imaged on a TS100 Eclipse inverted microscope (Nikon) equipped with a DS-1QM camera (Nikon). After that, HaCaT cells were treated with the peptide at a final concentration of 5 µM. Images were taken at 24, 48, and 72 h, and the area of opening was measured using ImageJ software. For each photograph, 10 measures of the open wound were recorded, averaged, and compared with the control samples. GraphPad Prism software was used for statistical analysis, performed by a Tukey test.
Groups of 20 wild-type (WT) zebrafish larvae (three days post-fertilization) were anesthetized with MS-222, submitted to a tail fin cut, and exposed to the peptide at a final concentration of 5 µM. All zebrafish larvae were imaged using a Nikon AZ100 lens equipped with a DS-Fi1 camera (Nikon). Measures of the tail fins at day 0, 2, 4, and 7 were performed, using the posterior notochord as a reference to the end of the tail.
Groups of 20 wild-type (WT) zebrafish larvae (three days post-fertilization) were anesthetized with MS-222, submitted to a tail fin cut, and exposed to the peptide at a final concentration of 5 µM. All zebrafish larvae were imaged using a Nikon AZ100 lens equipped with a DS-Fi1 camera (Nikon). Measures of the tail fins at day 0, 2, 4, and 7 were performed, using the posterior notochord as a reference to the end of the tail.