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. 2024 Jun 18;15:5199. doi: 10.1038/s41467-024-49597-z

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© The Author(s) 2024

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 Western blot analysis of GRP75 proteins expression in BV2 cells subjected to transfection of GRP75 siRNA or negative control (NC) siRNA (n = 3 batches of transfected cell samples; P = 0.9897; P = 0.0003). b Representative TEM images of ER-mitochondria contact (left panel) and the quantitative analysis of the distance (right panel, n = 3 mitochondria in 3 fields per condition; Interaction, F _{(1, 8)} = 19.14, P = 0.0024). Scale bar: 1 μm. c Representative fluorescence images (left panel) and quantitative analysis (right panel, n = 3 per condition; Interaction, F _{(1, 8)} = 11.11, P = 0.0103; Interaction, F _{(1, 8)} = 33.60, P = 0.0004) of Mag-Fluo-4 and Rhod-2 in BV2 cells. Scale bar: 20 μm. d Western blot analysis of PERK and elf2α phosphorylation proteins (n = 3 batches of transfected cell samples; Interaction, F _{(1, 8)} = 10.79, P = 0.0111; Interaction, F _{(1, 8)} = 124.7, P < 0.0001). e Representative confocal images (above panel) of the intracellular localization of lysosome (Lyso-Tracker, red), mitochondria (Mito-Tracker, green) and nucleus (Hoechst, blue) in BV2 cells and the statistical analysis (below panel, n = 4; Interaction, F _{(1, 8)} = 77.58, P < 0.0001) of the fluorescence intensity. Scale bar: 20 μm. f Representative photographs of DCF fluorescence (left panel) and Relative DCF fluorescence was quantitatively analyzed (right panel, n = 3; Interaction, F _{(1, 8)} = 4.353, P = 0.0704). Scale bar: 20 μm. g Representative images of MitoSOX (red) and Mito-tracker (green) fluorescence in BV2 cells (left panel). And relative MitoSOX fluorescence was quantitatively analyzed (right panel, Interaction, F _{(1, 8)} = 8.536, P = 0.0192). Scale bar: 20 μm. h Representative blots of co-immunoprecipitation (IP, left panel) and quantification of the NLRP3, Caspase-1 and ASC protein changes (n = 3 batches of transfected cell samples; Interaction, F _{(1, 8)} = 11.48, P = 0.0095; Interaction, F _{(1, 8)} = 140.9, P < 0.0001; Interaction, F _{(1, 8)} = 14.41, P = 0.0053). One-way or Two-way ANOVA with Tukey’s multiple comparisons test. Data are expressed as mean ± SEM. ns, p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001. Source data are provided as a Source Data file.