Validation of the Oncofetal (OnF) phenotypic reporter

To test its specificity, VAKSP tumoroids expressing the OnF reporter were sorted based on their GFP expression status. We then performed gene expression profiling analysis using data obtained from mRNA-seq of GFP high and GFP negative VAKPS cells. We first generated a VAKPS line stably expressing the OnF reporter. After selection, these tumoroids were expanded. Then GFP high and GFP negative cells were collected for RNA extraction using Fluorescence-Activated Cell Sorting.