The following plasmids were utilized: pMSCV-IRES GFP9 (link), pMXs-IRES-Puro (Cell Biolabs Inc), pMSCV-IRES-mCherry FP (a gift from Dario Vignali, Addgene #52114), pMSCV-loxp-dsRed-loxp-eGFP-Puro-WPRE (a gift from Hans Clevers, Addgene #32702). The CSF3RT618I mutation was generated previously9 (link). Full-length CEBPA and JAK3 cDNA was obtained from Genecopia. The CEBPA and JAK3 mutations were generated using the Quikchange Site Directed Mutagenesis Kit (Agilent) using the primers in Supplementary Table 1. To produce retrovirus, 293T17 cells were transfected with EcoPac helper plasmid (a gift from Dr. Rick Van Etten) and the appropriate transfer plasmid. Conditioned media was harvested 48–72 h after transduction.
Cdna CebpaCells Conditioned media Ires Jak3 Mutations Plasmid Primers Retrovirus Site directed mutagenesis
Corresponding Organization : Oregon Health & Science University
Other organizations :
Fred Hutch Cancer Center, MRC Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, University of Washington, Seattle University, Howard Hughes Medical Institute
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