scRNAseq of RPE-hTert cells expressing different levels of predictive reporter construct

Single cell sequencing of RPE1-hTert cells transduced with the NLS-mVenus-P2A-ER:HRAS^G12V construct, and then flow-sorted into subpopulations based on increasing mVenus intensity, corresponding to different levels of oncogenic RAS stabilised upon induction with 4OHT. Overall design: RPE1-hTert cells were infected with the predictive reporter (NLS-mVenus-P2A-ER:HRAS^G12V) construct then flow sorted into subpopulations based on increasing mVenus intensity ('S' < 'M' < 'L' < 'XL'). Cells were grown either in monoculture i.e. each subpopulation separately, or in coculture i.e. subpopulations mixed together in equal proportions, and induced with 4OHT. A total of 6 samples (uninduced coculture, induced coculture, induced 'S', induced 'M', induced 'L' and induced 'XL') were hashtagged with TotalSeq-A hashtag antibodies and pooled for encapsulation and single-cell sequencing using the 10x Genomics platform. Induced samples were harvested on Day 6 post-induction.