Fig. 1. Schematic representation of the MANO method.

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Fig. 1. Schematic representation of the MANO method. Schematic representation of the MANO method. Mouse 3T3 or Ba/F3 cells were infected with recombinant retrovirus expressing oncoproteins with corresponding 6-bp bar codes. Equal numbers of the stably transduced cells were mixed and cultured with different types of medium and/or treated with TKIs or vehicle. gDNA was harvested from the mixture of the remaining viable cells at the appropriate periods for each assay. The bar code sequences were PCR-amplified and subjected to deep sequencing on MiSeq sequencers to quantitate their relative abundance (a direct reflection of the cell number). To evaluate the transforming potential of each oncoprotein, the read number for each bar code was normalized to that of day 0. To evaluate the inhibition profile of test compounds across transduced clones in the mixture, the read number for each bar code was normalized to that of the vehicle-treated control. WT, wild-type. Shinji Kohsaka et al., Sci Transl Med 2017;9:eaan6566 Published by AAAS