First instar larvae were collected at 12 h after larval hatching and starved for 4 h. Brain lobes were dissociated from 40 to 45 starved larvae. Cells were dissociated with collagenase, filtered, spun, and resuspended in PBS.

Dmel R6.32

Reads were processed using Cell Ranger v2.2 (10x Genomics). They were mapped to the Drosophila melanogaster reference genome R6.22. Further analysis was performed using Seurat v3.0. For quality filtering, cells with less than 200 genes, more than 4,500 genes, or more than 20% of mitochondrial genes, were excluded. For dimensionality reduction, 31 principal components were selected visually on elbow plots as input for Uniform Approximation and Projection (UMAP) clustering.