Figure 1 | Scientific Reports

Figure 1

From: AUTS2 disruption causes neuronal differentiation defects in human cerebral organoids through hyperactivation of the WNT/β-catenin pathway

Figure 1

Generation and characterization of WT, AUTS2+/- and AUTS2-/- cerebral organoids. (A) The Integrative Genomics Viewer (IGV) snapshot of H3K4me3 ChIP-seq data from hESC and HEK293 cells shows two notable peaks in the region flanking exon 6 of AUTS2 (highlighted with a yellow box), suggesting the presence of intronic promoters. Red lines mark the sgRNA target region for creating AUTS2 knockout hESCs. The successful deletion of the targeted 190 kb region upstream of AUTS2 exon 6 is confirmed by Sanger sequencing results. (B) Immunoblot analysis confirmed the loss of AUTS2 expression in WT, AUTS2+/- , and AUTS2-/- hESC lines, with GAPDH serving as the loading control. For immunoblotting, at least three cerebral organoids for a total of three biological replicates were pooled (n = 3). (C) Microscopic examination reveals the morphology of 40-day-old cerebral organoids for each genotype. Ventricle-like structures in AUTS2-/- cerebral organoids are indicated with arrowheads. (D) The size of 21-day-old organoids was measured across genotypes: WT (n = 20), AUTS2+/- (n = 16), and AUTS2-/- (n = 22). Each bar in the graph represents the mean size, with error bars indicating the standard error. Student T-test was performed and statistical significance is marked with ** (P < 0.01).

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