Extended Data Fig. 9: Conservation analysis and lab mouse LacZ transgenic assays of Glider Accelerated Regions (GARs).

a, b, LacZ transgenic assays in E9.5 and E11.5 laboratory mouse embryos. Shown is the schematic of the construct and insertion site used to test GAR activity (a) and results from the assays (b). E11.5 embryos in (b) show expected neural tube staining from basal Shh promoter activity (black arrowheads). Scale bars: 500 µm (E9.5) and 1000 µm (E11.5). c, Conservation scores for genomic regions surrounding GARs and visualized against the reference sugar glider sequence as a heatmap (darker colours indicate higher conservation). Labelled in red are GARs that showed functional activity through luciferase assays, whereas GARs in black are those that did not. d, Opossum (Monodelphis domestica) genomic region containing the Emx2 locus and all Emx2-associated GARs. All Emx2-associated GARs have high-confidence orthologous matches with the opossum. Only the GAR located in the Emx2 promoter had a high-confidence orthologous match with eutherian genomes (Mus musculus and Homo sapiens). The orientation of this region in opossum is inverted with respect to the other species.