scRNA-seq data for H-23 and H-358 cells traced under using customized barcode

We barcoded clones with specific sequences and split the pools of cells into different sample that were treated with DMSO, single agent Sotorasib (AMG-510) or combination of Sotorasib and TEAD inhibitor (G'7883) for different duration. We performed this experiment for two cell lines (H-23 and H-358) independently. NCI-H358 and NCI-H23 cell lines were barcoded with TraCe-seq barcoding libraries and expanded in culture to establish the starting TraCe-seq cell populations. A fraction of the starting populations were profiled by single cell RNA-sequencing (scRNA-seq) to capture the relative clonal abundance and transcriptional profile of each barcoded clones right before drug treatment. In parallel, the rest of the cells were treated with sotorasib alone or in combination with GNE-7883 until resistant clones emerged under sotorasib single agent treatment. We conducted scRNA-seq profiling to capture acute responses to the treatment at 72h post treatment as well as after resistance has emerged to capture adaptive changes. These time points were chosen to allow us capture the differential transcriptional responses in the resistant versus sensitive clones, and compare the responses to sotorasib single agent versus sotorasib + GNE-7883 combination at individual clone and single cell level.