Fig. 2
Identification of candidate genes involved in cortical development via PB-based insertional mutagenesis. a The procedure used for the genetic screening. Cortical and ectopic layers containing cells electroporated with PB/PBase (red) at E14.5 were isolated surgically at P10. Genomic DNA (gDNA) was then extracted for splinkerette PCR to identify the insertion sites. b The products of the splinkerette PCR from four different mouse brains separated by agarose gel electrophoresis. Each prominent band (arrowheads) was excised for DNA sequencing. c Distribution of PB insertions within 50 kb of known genes in the mouse genome. Percentage of the PB insertions located in exons, introns, 5′ intergenic regions and 3′ intergenic regions are shown. d IPA was used for the functional categorization of the candidate genes. The area of each rectangle represents the p-value of a particular gene relevant to a particular functional pathway. The results show that these genes are mainly involved in development and nervous system functions. e A Circos diagram illustrating the positions of the identified candidate genes within the mouse genome and the type of insertion (intron and exon). Genes involved in three pathways were also linked together; these were genes involved in abnormal morphology of the brain, head and nervous system (yellow strings), genes involved in abnormalities of the craniofacial area, rhombencephalon and telencephalon (blue strings), and genes involved in cell movement of embryonic cells as well as cell–cell adhesion (red strings)
