Psn antibiotic mixture

U2OS cells were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). U2OS cells were cultured in DMEM (Gibco) supplemented with 10% FBS (Gibco) and 1% penicillin–streptomycin–neomycin (PSN) antibiotic mixture (Gibco) at 37 °C with 5% CO₂. To evaluate the transfection efficiency, FANCA plasmids were co-transfected with pEGFP-N2 vector (Clontech) into U2OS cells using Lipofectamine 3000 (Invitrogen) according to the manufacturer’s instructions. In experiments evaluating cell sensitivity to ICL damage, 2 μM mitomycin C (MMC; Sigma) was added into the culture medium 24 h after transfection.

Human embryonic kidney 293T (HEK293T) cells were purchased from the Cell Bank of the Chinese Academy of Sciences. HEK293T cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) (Gibco) supplemented with 10% fetal bovine serum (FBS) (Gibco) and 1% penicillin–streptomycin–neomycin (PSN) antibiotic mixture (Gibco) at 37℃ with 5% CO₂. HEK293T cells were transfected with the wild‐type or mutated WT1 plasmids using Lipofectamine 3000 (Invitrogen) according to the manufacturer's instructions.

Cell lines HCT116 and SW480 were from ATCC. SW620 and HT29 were from NCI. HCT116 p53 −/− was a kind gift from Dr. Bert Vogelstein. Caco2 and Colo 320 DM cell lines were procured from the national repository of National Centre for Cell Sciences, Pune, India. DAT1 was synthesized by slight modification of the method described earlier [63 (link)]. Vinblastine, taxol, 5-Fluoro uracil, DAPI, propidium iodide and U0126 were from Sigma, USA. siRNA construct for ERK1/2 was obtained from Santacruz Biotechnology. DMEM and PSN antibiotic mixture were purchased from Invitrogen, USA. MTT and Caspase 3 fluorogenic substrate were from USB and BD pharmingen, respectively. Tunnel assay kit was from R & D Biosystems, USA and immunohistochemical kit was purchased from Vector Labs, USA. Kits for detection of ALP, AST, ALT, Creatinine and BUN levels in serum were from Aspen Laboratories, India.