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. 2021 Dec 10;24(12):103484. doi: 10.1016/j.isci.2021.103484

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© 2021 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Senp5L/5S regulate neurite outgrowth and polarization

(A–C) Senp5 shRNAs (shRNA #01 or shRNA #02) or control shRNA was introduced into primary cultured cortical neurons along with a reporter plasmid encoding red fluorescent protein (RFP) (red). (A) Neurofilaments in individual neurons were stained with anti-SMI312 (green) at 3 div. (B) The stacked bar chart shows the percentages of cells having SMI312⁺ axons (stage 3). Numbers in parentheses indicate the number of cells analyzed. ∗∗∗, p < 0.001; chi-square tests with Holm-Bonferroni correction. (C) Box and whisker plots summarize the numbers of RFP⁺ neurites extending from individual neurons. Numbers in parentheses indicate the number of cells measured in three independent experiments. ns, not significant; ∗∗∗, p < 0.001; Wilcoxon rank sum tests with Holm-Bonferroni correction.

(D–F) Overexpression of Senp5L and Senp5S in immature neurons. pIRES2-mCherry-Senp5L, pIRES2-mCherry-Senp5S, or control pIRES2-mCherry was electroporated into primary cultured cortical neurons. Neurofilaments and mCherry were stained with anti-SMI312 (green) and anti-RFP (red) at 3 div. (E) The stacked bar chart shows the percentages of cells having SMI312⁺ axons (stage 3). Numbers in parentheses indicate the number of cells measured in three independent experiments; ∗∗∗, p < 0.001; chi-square tests with Holm-Bonferroni correction. (F) Box and whisker plots summarize the numbers of RFP⁺ neurites extending from individual neurons. Numbers in parentheses indicate the number of cells measured. ns, not significant; ∗∗∗, p < 0.001; Wilcoxon rank sum tests with Holm-Bonferroni correction. Scale bars, 20 μm.