Supplementary Figure 10: Human SETD5 rescue experiment.

a, Schematics of the localization of de novo SETD5 mutations in ID and ASD patients; each mutation is colour-coded based on the study reporting the de novo mutation; mutations used in this study are highlighted in bold. b-d, Representative example of the bulk enrichment sort and single cell sorting strategy. Shape analysis scatter plots (b). To exclude doublets, live cells are gated on forward scatter/ side scatter (FSC/SSC, left) followed by SSC (middle) and FSC (right). Scatter plots (c) of primary bulk enrichment for Setd5-P2A-GFP and CRISPR/Cas9-T2A-Cerulean3 positive cells (right) compared to WT non-transfected control (left). Scatter plots (d) showing enrichment for Setd5-P2A-GFP cells over the course of the secondary bulk enrichment sort (middle) and tertiary single cell sort (right). Auto-fluorescence was measured to ensure GFP positive cells.