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. 2023 Aug 8;30(9):2053–2065. doi: 10.1038/s41418-023-01203-3

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© The Author(s), under exclusive licence to ADMC Associazione Differenziamento e Morte Cellulare 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

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Fig. 5 — A Pd1 deletion results in global gene expression variation. Hierarchical clustering analysis of genes that change fold is more than two between Pd1^fl/fl mice and Pd1^cKO mice. B Gene ontology analysis of dysregulated genes in Pd1^cKO mice brains. C Image of Volcano plots shows the downregulated (green) and upregulated genes (red). D The mRNA level of Pd1 is obviously reduced upon Pd1 deletion. n = 4 individual experiments and p = 0.00001. E Loss of Pd1 results in an obvious increase in the expression of Pax3. n = 4 individual experiments and p = 0.002 for Pax3; p = 0.116 for Tlx3; p = 0.172 for Dgkg. F Downregulation of Pax3 leads to abnormal cell distribution in brain development. The plasmid of control and Pax3-sh1 were electroporated into the embryonic brain respectively at E13 and collected at E16 for further analysis. G Percentage of GFP⁺ cells in each zone of the cerebral cortex. n = 4 mice, p = 0.007 for VZ/SVZ; p = 0.468 for IZ; p = 0.002 for CP. H Abnormal cell distribution caused by Pd1 downregulation is partially rescued by Pax3-shRNA. I Graph shows the percentage of GFP⁺ cells in each zone. p = 4 mice, p = 0.009 (Control&Pd1-sh1) and p = 0.889 (Control&Pd1-sh1+Pax3-sh1) for VZ/SVZ; p = 0.0003 (Control&Pd1-sh1) and p = 0.726 (Control&Pd1-sh1+Pax3-sh1) for IZ; p = 0.0005 (Control&Pd1-sh1) and p = 0.753 (Control&Pd1-sh1+Pax3-sh1) for CP. J BrdU incorporation defects induced by Pd1 knockdown are rescued by Pax3 suppression. K Graph shows the percentage of BrdU incorporation in GFP⁺ cells in VZ/SVZ. n = 5 mice, p = 0.014 for Control&Pd1-sh1; p = 0.383 for Control&Pd1-sh1+Pax3-sh1. L Downregulation of Pax3 overrides neuronal differentiation defects caused by Pd1 knockdown. M Graph shows the percentage of Tuj1⁺GFP⁺ cells relative to all GFP⁺ cells. n = 5 mice, p = 0.002 for Control&Pd1-sh1; p = 0.884 for Control&Pd1-sh1+Pax3-sh1. Error bars represent means ± S.E.M.; Two-tailed unpaired t-test, P < 0.05(*), P < 0.01(**), n.s. not significant. The scale bar represents 100 μm.