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. 2022 Jul 28;19:194. doi: 10.1186/s12974-022-02557-0

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© The Author(s) 2022

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. 7 — Activation of ERK1/2 and NF-κB pathways by PMA partially reversed the anti-pyroptotic effect of TGF-β1 in vitro. A The protein expression of Cleaved-GSDMD, NLRP3, and IL-1β in cultured microglia under different treatments was evaluated by Western blot. B Quantitative analysis of Western blot was performed using One-way ANOVA followed by Tukey’s multiple comparisons test. **P < 0.01, ****P < 0.0001 versus CT group; #P < 0.05, ##P < 0.01 versus LPS group; &P < 0.05, &&P < 0.01 versus LPS + TGF-β1 group. N = 6 per group. C Representative confocal images (scale bar = 30 μm) and 3D reconstructed images in high power field (scale bar = 10 μm) of Iba1 and NLRP3 in cultured microglia under different treatments. D Quantitative analysis of the volume of NLRP3⁺ punctures per cell in immunofluorescent staining. One-way ANOVA followed by Tukey’s multiple comparisons test was used for statistical analysis. ****P < 0.0001 versus CT group; # P < 0.05 versus LPS group, &P < 0.05 versus LPS + TGF-β1 group. N = 9 per group. E The level of IL-1β in microglial CM under different treatments was analyzed by ELISA. Quantitative analysis was performed using One-way ANOVA followed by Tukey’s multiple comparisons test. ****P < 0.0001 versus CT group; ###P < 0.001 versus LPS group; &P < 0.05 versus LPS + TGF-β1 group. N = 6 per group. F The level of TNF-α and IL-6 in microglia-CM exposed to varied stimulations was measured by CBA. Quantitative analysis was performed using One-way ANOVA followed by Tukey’s multiple comparisons test. ****P < 0.0001 versus CT group; ###P < 0.001, ####P < 0.0001 versus LPS group; &P < 0.05, n.s. no significance versus LPS + TGF-β1 group. N = 6 per group. G Representative images of bright field (scale bar = 40 μm) and immunostaining with Iba1 (scale bar = 30 μm) in cultured microglia exposed to varied treatments. H Quantitative analysis of the percent of amoeboid-like microglia in bright-field images. ****P < 0.0001 versus CT group; ##P < 0.01 versus LPS group; &P < 0.05 versus LPS + TGF-β1 group. N = 6 per group