Rpmi 7951 htb 66

Dithiothreitol (DTT), iodoacetamide, ammonium bicarbonate (Ambic), ammonium hydroxide, sodium docecylsulphate (SDS), Trifluoroacetic acid (TFA), sodium deoxycholate (SDC), tris(hydroxymethyl)aminomethane (Tris), formic acid, and urea were purchased from Sigma–Aldrich (St. Louis, MO, USA). Triethylamonium bicarbonate (TEAB) and hydroxylamine were from Thermo Fisher Scientific. Water and organic solvents were all LC–MS grade and supplied by Merck (Darmstadt, Germany) or (Thermo Fisher Scientific). Endoproteinase Lys‐C was obtained from Wako (Osaka, Japan) and sequencing‐grade modified trypsin was purchased from Promega (Madison, WI, USA). Cell lines SK MEL2 (HTB‐68), SK MEL28 (HTB‐72), and RPMI‐7951 (HTB‐66) were obtained from the American Type Culture Collection (ATCC).

Human malignant melanoma cells A375 (CLR-1619) and RPMI-7951 (HTB-66) were obtained from American Type Culture Collection (Manassas, VA, USA). SK-MEL-28 cells were obtained from Alan Houghton, Sloan-Kettering Institute for Cancer Research (New York, NY, USA). A375 and SK-MEL-28 cells were maintained in DMEM (Mediatech Inc., Manassas, VA, USA) and RPMI-1640 (HyClone Laboratories, Logan, Utah, USA) medium respectively supplemented with 10% heat-inactivated fetal bovine serum (Sigma-Aldrich Corporation, St. Louis, MO, USA) and 100 mg/ml penicillin-streptomycin (Mediatech Inc., Manassas, VA, USA). RPMI-7951 cells were cultured in EMEM medium (Quality Biologicals Inc., Gaithersburg, MD) supplemented with 10% heat-inactivated fetal bovine serum and 100 mg/ml penicillin-streptomycin. Cells were maintained at 37°C and 5% CO₂ in a 95% humid environment. Fisetin and sorafenib used for the treatment of the cells were dissolved in DMSO (Fischer Scientific, Fair Lawn, NJ, USA). The final concentration of DMSO was ≤ 0.1% (v/v) in each treatment.

Dithiothreitol (DTT), iodoacetamide, ammonium bicarbonate (Ambic), ammonium hydroxide, sodium docecylsulphate (SDS), trifluoroacetic acid (TFA), sodium deoxycholate (SDC), tris(hydroxymethyl)aminomethane (Tris), formic‐acid, and urea were purchased from Sigma‐Aldrich (St. Louis, MO, USA). Triethylamonium bicarbonate (TEAB) and hydroxylamine were from Thermo Fisher Scientific. Water and organic solvents were all LC‐MS grade and supplied by Merck (Darmstadt, Germany) or (Thermo Fisher Scientific). Endoproteinase Lys‐C was obtained from Wako (Osaka, Japan) and sequencing‐grade modified trypsin was purchased from Promega (Madison, WI, USA). Cell lines SK MEL2 (HTB‐68), SK MEL28 (HTB‐72), and RPMI‐7951 (HTB‐66) were obtained from the American Type Culture Collection (ATCC).