Sk mel 2

All cell lines were purchased from the American Type Culture Collection (ATCC) and maintained at 37 °C with 5% CO₂. HEK293T (ATCC®CRL-3216), A375 (ATCC®CRL-1619), G361 (ATCC®CRL-1424), MeWo (ATCC®HTB-65), SK-MEL-5 (ATCC®HTB-70), SK-MEL-2 (ATCC®HTB-68), B16 (ATCC®CRL-6475), and HT29 (ATCC®HTB-38) cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (Gibco, 10437028), 2 mM l-glutamine, and 1% penicillin–streptomycin (Invitrogen, 15140122). Transfections were performed using Calcium Phosphate Transfection Kits (Clontech, 631312) or PolyFect Reagent (Qiagen, 301107), following the manufacturer’s instructions. None of the cell lines used in this study was found in the database of commonly misidentified cell lines that is maintained by ICLAC and NCBI Biosample. All cell lines were tested and confirmed to be free of mycoplasma.

Human melanoma cell lines CHL-1 and SK-MEL-2 were obtained from Cell Resource Center, Chinese Academy of Sciences. CHL-1 and SK-MEL-2 cells were cultured in DMEM (Gibco) and MEM (Gibco) medium, respectively, supplemented with 10% fetal bovine serum (Gibco) at 37 °C with 5% CO₂. Cell lines used in this study were authenticated by STR profiling and routinely tested as mycoplasma-free.

All cell lines were purchased from ATCC and have been authenticated and exponentially grown at 37 °C and 5% CO₂. HeLa and HEK293 were cultured in DMEM (Gibco, MA USA) supplemented with 10% FBS (Gibco) and 2 mM l-Glutamine (Gibco). A375, SK-Mel2, SK-Mel28, SK-Mel239 were cultured in RPMI (Gibco) supplemented with 10% FBS (Gibco) and 2 mM l-Glutamine (Gibco). Vemurafenib (PLX4032) and Birinapant were purchased from Selleck Chemicals (TX, USA).

Human cells from Lung Squamous Cell Carcinoma (SK-MES-1), Glioblastoma Multiforme (U251), Uterine Corpus Endometrial Carcinoma (HEC-1B), and Skin Cutaneous Melanoma (SK-MEL-2) were from the American Type Culture Collection (ATCC). SK-MES-1 and U251 cells were cultured in DMEM (GIBCO, USA) supplemented with 10% fetal bovine serum (FBS, GIBCO, USA). HEC-1B cells were incubated in Roswell Park MEMorial Institute-1640 (RPMI-1640, GIBCO, USA) supplemented with 10% FBS. SK-MEL-2 was cultured in MEM (GIBCO, USA) supplemented with 10% FBS. All the cells were cultured in a humidified incubator at 37°C and a 5% CO₂ atmosphere. Cells were harvested in logarithmic growth phase in all the experiments performed in this study.

Human normal melanin cell line, PIG1 (American type culture collection, Manzas, Virginia, USA), and malignant melanoma cells, A375 and SK-MEL-2 cells (iCell Bioscience Inc, Shanghai, China) were maintained in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal bovine serum at 37° C with 5% CO₂. The pcDNA3.1-ALDH2 plasmid synthesized by GenePharma (Shanghai, China) was transfected into A375 and SK-MEL-2 cells with the presence of Lipo3000 transfection reagent (Thermo Fisher Scientific, Waltham, MA, USA). Transfected cells were harvested after 48 h of incubation in 6-well plates and the transfection efficiency was measured using quantitative real-time polymerase chain reaction (qRT-PCR).