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. 2023 Mar 29;14(3):227. doi: 10.1038/s41419-023-05660-z

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© The Author(s) 2023

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. 6 — The BV2 murine microglial cell lines were treated with high glucose (50 mM) for 48 h. For treatments, cells were pre-stimulated with GSK-872 (5 μM) before high glucose stimulation for 12 h. D-Mannitol was used as an osmotic control. A The protein levels of RIP1 and RIP3 were detected by western blotting. B The protein levels of MLKL and p-MLKL were detected by western blotting. C, D Combined p-MLKL immunostaining and TUNEL assays on BV2 microglial cells under high glucose and normal condition. Ten images in each group were randomly chosen for analysis. E, F The levels of TNF-α and MCP-1 in cultured microglia was detected using TNF-α and MCP-1 ELISA kit respectively. HG high glucose. GSK, the specific RIP3 inhibitor GSK-872. Western blotting and ELISA assays were repeated 3 times. Data were mean ± S.D. *P < 0.05, **P < 0.01, ***P < 0.001, ns no significance.