Fig. 1

Detection and validation of circRNAs in human neuronal differentiation. a Schematic representation of sample and data collection. b Venn diagram of circRNAs detected from three different circRNA detection programmes, numbers highlighted in bold indicate circRNAs included in downstream analysis. c Volcano plots of differentially expressed circRNAs at D5 and D28 of differentiation compared to NES. d Venn diagram of significantly dysregulated circRNAs (padj < 0.05 and LFC ≥|2|). e Cluster analysis of circRNAs differentially expressed throughout differentiation. f Detection of top differentially expressed circRNAs by PCR with divergent junction primers from RNAse R-treated samples. Samples from D28 were used to detect 15 increased circRNAs (left) and samples from NES cells were used for detection of 5 decreased circRNAs (right). Asterisks indicate reactions with additional bands or no product, blue highlighting indicates the product is an isoform of the intended circRNA target, and red highlighting indicates PCR artefact. g Example schematic of Sanger sequencing validation of circRNA exon retention in circATRNL1. Exons are shown in purple, green arrows indicate primer location, and sequencing alignment is shown in orange