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. 2018 Mar 29;103(7):1169–1181. doi: 10.3324/haematol.2017.183970

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Figure 1. — Large-scale shRNA-based RNAi screens identify novel target genes required for cell growth of AML cells. A. Schematic illustration of the shRNA-based screening procedure. B. Bar chart representing ratios of barcodes from 27104 individual shRNAs after screens in murine MLL-AF9 transformed AML cells compared to murine FDCP-mix BMs, after 10 cell divisions (T10). The 1086 bar-codes with at least a five-fold lower representation in the murine MLL-AF9 AML cells compared to the murine FDCP-mix cells are highlighted in red. C and D. Bar charts representing differences in ratios of barcodes from 27491 individual shRNAs (THP-1) and 27104 (NOMO-1) after pooled loss of function screens after T10. Individual shRNAs targeting CHD4 are highlighted in red. E. Venn diagram illustrating the overlap between target genes that are five-fold more depleted in mouse AML cells compared to mouse FDCP-mix control cells, five-fold depleted target genes in THP-1 cells and NOMO-1 cells after T10. F. List of the 34 target genes overlapping in all three screens in the Venn diagram from Figure 1E together with information about the number of individual shRNAs resulting in at least a five-fold reduction in abundance of barcodes in the screens of the human AML and the mean fold reduction in abundance of barcodes in the screens after T10. FDCP: factor dependent cell-Paterson; AML: acute myeloid leukemia; PCR: polymerase chain reaction; shRNA: short hairpin RNA.