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. 2021 Jul 5;12:4113. doi: 10.1038/s41467-021-24376-2

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Immunoprecipitation by α-TubK40me3 antibody and following immunoblotting with α-tubulin showed that the level of α-TubK40me3 was increased in HEK293 cells transfected with Flag-SETD2(1469-1724) and Flag-NES-SETD2(1469-1724), while the level of H3K36me3 was not significantly changed (n = 3 biological replicates). b Immunostaining of Flag (green) and α-tubulin (red) showed the subcellular localization of Flag-SETD2(1469-1724) and Flag-NES-SETD2(1469-1724) in HEK293 cells (n = 3 biological replicates). Cells were stained for DAPI (blue). Scale bar: 5 μm. c Representative images of coronal brain sections in somatosensory cortex at E16 showed the distribution of GFP⁺ cells (green) electroporated with GFP reporter as well as shNC, mSETD2 shRNA2 or mSETD2 shRNA2 together with Flag-NES-SETD2(1469-1724) at E14. Sections were stained for DAPI (blue). Scale bar: 25 μm. d Quantitative analysis of c showed that the migration defects induced by SETD2 knockdown was rescued by expressing Flag-NES-SETD2(1469-1724) (n = 6, 12, 13, respectively). e Representative images of GFP⁺ neurons (green) in IZ. Brain slices were stained for DAPI (blue). Scale bar: 10 μm. f Quantitative analysis of e showed that the percentage of neurons at the multipolar stage in IZ was rescued by expressing Flag-NES-SETD2(1469-1724) (n = 6, 12, 13, respectively). g Representative images of coronal brain sections in somatosensory cortex at E16 showed the distribution of GFP⁺ cells (green) electroporated with GFP reporter as well as shNC, mSETD2 shRNA2 or mSETD2 shRNA2 together with Flag-α-tubulin^K40F/K40A at E14. Sections were stained for DAPI (blue). Scale bar: 25 μm. h Quantitative analysis of g showed that the migration defects induced by SETD2 knockdown was rescued by expressing α-tubulin^K40F but not α-tubulin^K40A (n = 28, 22, 10, 12, respectively). i Representative images of GFP⁺ neurons (green) in IZ. Brain slices were stained for DAPI (blue). Scale bar: 10 μm. j Quantitative analysis of i showed that the percentage of neurons at the multipolar stage in IZ was rescued by expressing α-tubulin^K40F but not α-tubulin^K40A (n = 28, 22, 10, 12, respectively). All data were shown as the mean ± s.e.m. and analyzed using two-way ANOVA with Bonferroni’s post-hoc test. *P < 0.05, ***P < 0.001 versus shNC; ^#P < 0.05, ^###P < 0.001 versus shRNA2. Source data are provided as a Source Data file.