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. 2020 Feb 18;11(2):125. doi: 10.1038/s41419-020-2324-4

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© The Author(s) 2020

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 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/.

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Fig. 3 — a, b Lentiviral vectors (SESN2 knockdown and SESN2 over-expression) were transduced DC2.4 cells, and scramble cells were transduced with blank vector. Transduced cells were analyzed by fluorescence microscopy (×400), scale bar = 200 μm, and then protein levels of SESN2 were determined by Western blotting. DC2.4 cells underwent SESN2-knockdown or SESN2-overexpression were incubated with 100 ng/ml HMGB1 for 48 h, and DC2.4 cells underwent blank vector treated with 100 ng/ml HMGB1 for 48 h served as the scramble group. Apoptotic rates of cells were measured with PE-Annexin-7-AAD by flow cytometry (c, d), and cell apoptosis was analyzed by Hoechst 33342 dye (blue) to observe nuclear condensation (×400, e, f), scale bar = 200 μm. Levels of SESN2, cleaved-caspase-3, cleaved-caspase-9, Bcl-2, and Bax were assessed by Western blotting (g, h), respectively. β-actin served as the internal standard. Values were represented as mean ± SD obtained from three independent experiments, n = 3 per group. Statistical significance: *P < 0.05 versus the scramble group; ^#P < 0.05 versus the scramble group treated with HMGB1.

Fig. 3 — a, b Lentiviral vectors (SESN2 knockdown and SESN2 over-expression) were transduced DC2.4 cells, and scramble cells were transduced with blank vector. Transduced cells were analyzed by fluorescence microscopy (×400), scale bar = 200 μm, and then protein levels of SESN2 were determined by Western blotting. DC2.4 cells underwent SESN2-knockdown or SESN2-overexpression were incubated with 100 ng/ml HMGB1 for 48 h, and DC2.4 cells underwent blank vector treated with 100 ng/ml HMGB1 for 48 h served as the scramble group. Apoptotic rates of cells were measured with PE-Annexin-7-AAD by flow cytometry (c, d), and cell apoptosis was analyzed by Hoechst 33342 dye (blue) to observe nuclear condensation (×400, e, f), scale bar = 200 μm. Levels of SESN2, cleaved-caspase-3, cleaved-caspase-9, Bcl-2, and Bax were assessed by Western blotting (g, h), respectively. β-actin served as the internal standard. Values were represented as mean ± SD obtained from three independent experiments, n = 3 per group. Statistical significance: *P < 0.05 versus the scramble group; ^#P < 0.05 versus the scramble group treated with HMGB1.