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. 2021 Feb 5;49(4):1935–1950. doi: 10.1093/nar/gkab030

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© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 7. — PAX6 interacts with NSD1 to regulate the H3K36me3 modification level and expression of the target neural genes. (A) Enrichment of H3K36me2 on SOX1 (TSS +1300), POU3F3 (TSS +1000), MSX1 (TSS +1000) and NR2F1 (TSS +1500) genomic region in control, PAU-PAKI and PAX6KO cells on day 16 of dorsal neural differentiation. (B) Co-IP analysis showing that HA-PAX6a interacts with myc-NSD1 in 293FT extracts, but not NSD2, NSD3, SETMAR, SETD3 or SMYD2. (C) Co-IP analysis showing that PAX6 interacts with myc-NSD1 in control and PAU-PAKI cells on day 16 of dorsal neural differentiation. (D) Enrichment of myc-NSD1 on SOX1 (TSS +1300), POU3F3 (TSS +1000), MSX1 (TSS +1000) and NR2F1 (TSS +1500) genomic region in control, PAU-PAKI and PAX6KO cells on day 16 of dorsal neural differentiation. (E) Co-IP analysis showing that myc-NSD1 interacts with Flag-PAX6a, Flag-ΔPD and Flag-ΔHD mutant in 293FT extracts, but not Flag-ΔC mutant. (F, G) Immunostaining assay of TBR1 (green) after PAX6a or ΔC mutant overexpression in the PAX6KO cells on day 25 of dorsal neural differentiation (F), and quantification of TBR1⁺ cells (G). Scale bar, 100 μm. (H) Summary of the PAUPAR/PAX6/NSD1-mediated transcriptional regulatory model. Data are presented as mean ± SEM. *P< 0.05, **P< 0.01 versus CTRL group (A, D and G); ^##P< 0.01 versus PAX6KO + ptPAX6a-dox group (G) (t-test/ANOVA). See also Supplementary Figure S7.