B16F10 cells were seeded in 6-well plate at a density of 5 × 104 cells/well and incubated for 16 h. They were then treated with various concentrations of peptide mixture in 2% FBS containing media for 3 days. To induce melanin production, 100 ng/ml α-MSH was co-treated and 200 μM arbutin was used as a positive control. RNA was isolated from cells using RNA extraction kit (Qiagen, Germany), and cDNA synthesis was performed using RT-PCR premix (iNtRON Biotechnology, Seongnam, Korea). After preparation of reaction mixture with PCR premix (iNtRON Biotechnology) and primers for each gene, PCR was performed using PCR machine (Eppendorf, Germany). Subsequently, mRNA expression patterns were determined by agarose gel electrophoresis.

HaCaT cells were seeded in 6-well plate at a density of 3 × 105 cells/well and incubated for 16 h. They were then treated with various concentrations of peptide mixture in the serum-free media for 1 h and additionally treated with 4U trypsin for 16 h. The cells were collected and RT-PCR was performed in the same manner as described above.

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