Fig 2: Knockdown of HSPA8 suppresses NF-?B and NLRP3 inflammasome activation in OGD/R-induced primary astrocytes. A Immunofluorescence of GFAP in primary rat spinal cord astrocytes was performed. Scale bar = 100 µm. B The relative protein level of HSPA8 in primary astrocytes was detected after injection of LV-sh-HSPA8 via western blot assay. C The relative protein level of total NF-?B P65, p-NF-?B P65, NLRP3, ASC, and p20/pro-caspase-1 in primary astrocytes was detected after injection of LV-sh-HSPA8 via western blot assay. D The contents of IL-1ß and IL-18 in primary astrocytes were detected by ELISA assay. E Relative protein levels of IL-1ß and IL-18 in primary astrocytes were detected via western blot assay. F The intracellular location of the NF-?B P65 was observed by immunofluorescence staining. Scale bar = 50 µm. G Quantification of NF-?B P65-immunofluorescence intensity in the nuclear was analyzed. H The DNA-binding activity of NF-?B in primary astrocytes was revealed by EMSA. Data were shown as means ± SD of three independent experiments. N = 3 cells per group. Statistical analysis was performed by one-way ANOVA followed by Turkey’s for multiple comparisons. *p < 0.05 vs. sham. # p < 0.05 vs. SCII + LV-sh-NC. NLRP3, nod-like receptor pyrin domain-containing 3; ASC, the apoptosis-associated speck-like protein containing CARD; SCII, spinal cord ischemia–reperfusion injury; IL-1ß/18, interleukin-1ß/18; NF-?B, nuclear factor-kappa B

Fig 3: DSF inhibits the formation of N-GSDMD and the release of IL-18 and IL-1ß in the lung tissue of PH mice. A Representative WB images and analysis of caspase-1 and CI. caspase-1 and IL-1ß (n = 5 each). B Representative WB images and analysis of GSDMD, N-GSDMD and IL-18 (n = 5 each). C Representative WB images and analysis of NLRP3 and ASC. D, E ELISA detection of inflammatory factors (IL-18 and IL-1ß) in the plasma, (n = 6 each). Values are the mean ± SEM. Statistical significance was analysed by one-way ANOVA followed by Bonferroni’s multiple comparison test or the Kruskal–Wallis test. *P < 0.05 vs. vehicle, and #P < 0.05 vs. normoxia

Fig 4: AIP1 expression is markedly reduced after alkali burn injury. A Representative slit-lamp images of the area of corneal neovascularization at different times after alkali burn injury (magnification: × 40). B Corneal opacity, neovessel size, and vessel size scores increased gradually from the third day of alkali burn injury and continued thereafter (N = nine). C Immunofluorescence staining revealed an obvious decrease in the expression of AIP1 and a concomitant increase in the expression of VEGFa after alkali burn injury (scale bar: 50 µm, magnification: × 400). D Western blot analysis demonstrating significantly upregulated expression of NOX4, NLRP3, ASC, clv-casp-1 and clv-IL-1ß and downregulated expression of AIP1 and NLRP6 in the cornea after alkali burn injury (N = three). The error bars represent the means ± SD, and comparisons were made using one-way ANOVA. clv-casp1, cleaved caspase-1; clv-IL-1ß, cleaved-IL-1ß. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001

Fig 5: PSG suppressed pyroptosis of the liver tissues in IBD mice. Effect of PSG on the levels of IL-1ß (a) and IL-18 (b) in the liver tissues of IBD mice. (c) Protein expression of NLRP3 in the liver tissues was detected by IHC. Protein expressions of NLRP3, gasdermin D, caspase 1, and ASC in the liver tissues were determined by Western blot assay (d) and quantitative analysis for Western blot results (n = 3) (e). Results were expressed as the mean ± SEM. *p < 0.05**p < 0.01 vs. the control group; #p < 0.05 and ##p < 0.01 vs. the model group (DSS only).