The human neuroblastoma cell lines SK-N-BE(1) and SK-N-BE(2) were obtained from Children’s Oncology Group (COG, USA). SK-N-BE(1) cell line was isolated from bone marrow of a 2-year-old patient when he was primary diagnosed without any treatment, and SK-N-BE(2) cell line was gained from the same patient when he has been received several courses of chemotherapy with cyclophosphamide, doxorubicin, as well as vincristine. These two cell lines had different drug sensitivity (Additional file 1 : Figure S1) [11 (link)–13 (link)].
SK-N-BE (1) and SK-N-BE (2) cell lines were maintained in RPMI 1640 (Gibco, USA) with 10% fetal bovine serum (Gibco, USA), 50 units/ml penicillin, 50 μg/ml streptomycin (Yeasen, China) and 1X ITS (Sodium Pyruvate 0.11 g/L, L-glutamine 1.5 g/L, NaHCO3 1.5 g/L (Yeasen, China)). These two cell lines were incubated humidified air supplemented 5% CO2 at 37 °C.
To enable the quantitative proteomics, we used specialized stable isotope labeling with amino acids in cell culture (SILAC) medium supplemented with 10% dialyzed fetal bovine serum (Invitrogen, Darmstadt, Germany) to label the cells, where deficient Arginine and Lysine were supplemented with either stable isotope encoded heavy Arginine and Lysine (Euriso-top) or normal Arginine and Lysine for the light. Labeling efficiency was confirmed before the following quantitative full proteome analysis.
SK-N-BE (1) and SK-N-BE (2) cell lines were maintained in RPMI 1640 (Gibco, USA) with 10% fetal bovine serum (Gibco, USA), 50 units/ml penicillin, 50 μg/ml streptomycin (Yeasen, China) and 1X ITS (Sodium Pyruvate 0.11 g/L, L-glutamine 1.5 g/L, NaHCO3 1.5 g/L (Yeasen, China)). These two cell lines were incubated humidified air supplemented 5% CO2 at 37 °C.
To enable the quantitative proteomics, we used specialized stable isotope labeling with amino acids in cell culture (SILAC) medium supplemented with 10% dialyzed fetal bovine serum (Invitrogen, Darmstadt, Germany) to label the cells, where deficient Arginine and Lysine were supplemented with either stable isotope encoded heavy Arginine and Lysine (Euriso-top) or normal Arginine and Lysine for the light. Labeling efficiency was confirmed before the following quantitative full proteome analysis.
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