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. 2020 Oct 28;17:321. doi: 10.1186/s12974-020-02001-1

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Irradiation induces microglial senescence in vitro. a Cytochemical staining for SA-β-gal in primary microglia and BV2 cells at 24 h after 10 Gy linac irradiation. Representative images are shown (scale bar, 100 μm). b The ratio of SA-β-gal positive cells is shown as the mean ± SEM; ***P < 0.001, versus the control group. c Primary microglia were exposed to a single dose of IR (10 Gy) by linac. Cells were immunostained with anti-Iba-1 (red) and anti-P16^INK4A (green) antibodies 24 h after IR. Representative images are shown (scale bar, 100 μm). d The ratio of P16^INK4A positive primary microglia is shown as the mean ± SEM; ***P < 0.001, versus the control group. e BV2 cells were exposed to a single dose of IR (10 Gy) by linac. Cells were immunostained with anti-P16^INK4A antibody (red) 24 h after IR. Representative images are shown (scale bar, 100 μm). f The ratio of P16^INK4A positive BV2 cells is shown as the mean ± SEM; ***P < 0.001, versus the control group. g Expression levels of Mmp3, Cdkn2a (P16^INK4A), Cdkn1a (P21), and p53 in BV2 cells at different time points after 10 Gy IR by linac were examined using qRT-PCR (mean ± SEM, ***P < 0.001). h The protein levels of p53 from BV2 cells and HT22 cells after exposure to different doses at different time points, as examined using western blotting. i Histogram of the quantification of the immunoreactive protein bands on the western blots (mean ± SEM, ns, not significant, ***P < 0.001)