Generation of the dnFOS plasmid was previously described16 (link) - dnFOS was amplified from cDNA provided by Charles Vinson33 (link) with SalI and NotI restriction site overhangs. Using these restriction sites, the fragment was ligated into pENTR2B (Addgene) and then recombined into pCW57.1 (Addgene). The empty vector was pCW57.1 alone. The shRUNX1::ETO plasmid was generated with XhoI and EcoRI restriction site overhangs. Using these restriction sites, the fragment was ligated into tRMPVIR (Addgene) plasmid. The shRNA sequence is 5′-AAACCTCGAAATCGTACTGAGA-3′. Plasmids were selected and propagated in DH5α competent cells prior to maxiprep using NucleoBond Xtra Midi EF kit and then lentiviral production. All unique biological materials are available from the authors upon request.
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