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. 2022 Apr 11;13(4):329. doi: 10.1038/s41419-022-04792-y

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© The Author(s) 2022

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. 1 — A Stable cells expressing short hairpin RNA against HuD (shHuD) or control RNA (shCtrl) were established using mouse neuroblastoma Neuro2A (N2a) cells. Downregulation of HuD was assessed via western blotting analysis. β-ACTIN was used as a loading control. B Cells were stained with senescence-associated β-galactosidase (SA β-gal). The relative intensity of SA β-gal was assessed via densitometric analysis using Image J. C The level of p16^INK4a was investigated via immunofluorescence microscopy and quantified by counting the number of p16^INK4a-expressing cells. Green: p16^INK4a, blue: nuclei. D The level of LAMIN B was assessed via western blotting analysis. β-ACTIN used as a loading control. E Cells were incubated with DCFDA, a fluorescent indicator of ROS and cellular ROS levels between N2a_shCtrl and N2a_shHuD cells were determined using flow cytometry. Images are representative, and data indicate the mean ± SEM from three independent analyses. Scale bar, 50 μm. The statistical significance of the data was analyzed via Student’s t-test; **p < 0.01.