Engineering an inducible leukemia-associated transcription factor enables large-scale ex vivo production of functional human phagocytes - Nanostring data

RNA expression analysis via NanoString nCounter Analysis System with four different cell types (three biological replicates each) using the myeloid innate immunity panel. Total RNA was isolated with the RNeasy Plus Mini Kit (Qiagen). The four different cell types include:
- Untransduced hematopoietic stem and progenitor cells (HSPCs) from healthy donors (cultivated for 3 days in media with IL-3, IL-6, SCF, TPO, GM-CSF, FLT3-L)
- HSPCs transduced with a retroviral construct DD-MLL-ENL-ires-GFP. DD is a destabilization domain leading to rapid degradation of MLL-ENL. The small molecule Shield-1 prevents degradation. The used cells were cultivated in the presence of Shield-1 for over 50 days to obtain an MLL-ENL expressing, GFP+ progenitor cell population. Cultivation in media with IL-3, IL-6, SCF, TPO, GM-CSF, FLT3-L.
- Macrophages differentiated from the MLL-ENL expressing progenitor cells after withdrawal of Shield-1 and cultivation in media with GM-CSF, LPS and IFNγ for 14 days.
- CD14+ monocytes derived from peripheral blood from healthy donors. PBMCs were isolated via density gradient centrifugation. CD14+ monocytes were isolated using the classical monocyte isolation kit from Miltenyi.

bulk RNAseq dataset MLL ENL hemCellcomp : 

Three independent replicates of MLL-ENL cells at progenitor and macrophage stages were used for transcriptomic profiling by RNA-sequencing. Data was compared to RNAseq data from various hematopoietic cells from published studies.