Brains from stage 17 embryos were dissociated and 24ALH larval brains were dissected and dissociated. These samples were kept separate and NSCs were enriched for by sorting for GFP+ cells via flow cytometry (using wor-GAL4>UAS-mCD8-GFP). Cells isolated by FACS were processed using the 10X Genomics Chromium Platform, using v3 chemistry (3' end gene expression), with the exception of Embryo_St17_biolrep1, which used v2 chemistry.