BCL2 targeting overcomes the microenvironment-induced resistance to crizotinib in ALK+ anaplastic large cell lymphoma
BCL2 targeting overcomes the microenvironment-induced resistance to crizotinib in ALK+ anaplastic large cell lymphoma
cell lines were from DSMZ (SUPM2, L82). MS-5 cells were kindly shared by Prof. Lucio Pastore (Department of Molecular Medicine and Medical Biotechnology, University of Napoli, Italy).  All cells were cultivated at 37°C in a humidified 5% CO2 atmosphere and were routinely tested for mycoplasma contamination using a Mycoplasma PCR detection kit (Applied Biological Materials Inc. abm # G238) and always tested negative. Drugs were purchased at MedChem express. All drugs were dissolved according to manufacturer's instructions. Total RNA extraction from cells was performed using TRIZOL (Invitrogen) according to the manufacturer's protocols.  Total RNA concentrations were determined by using NanoDrop spectrophotometer (Thermo Fisher) and their quality was assessed with the TapeStation 4150 (Agilent Technologies). After GEM generation, all the steps of cDNA synthesis, clean-up and amplification, DNA fragmentation, end-repair, adapter ligation and indexing PCR-amplification were done with the Chromium Single Cell 3' reagents following the manufacturer's protocol while the DNA purification as well as the size selection steps were done with SPRIselect magnetic beads (Beckman Coulter, Brea, CA) as indicated by the 10X protocol. Finally, the concentration of each indexed library was determined by using Qubit dsDNA BR Assay (Thermo Fisher Scientific, USA) and their quality were assessed with the D1000 screentape on the TapeStation 4150 (Agilent Technologies, Santa Clara, CA).