Fig 1: NLRP3 Promotes Pyroptosis of Human NP Cells Stimulated by Lactate. (A-C) ASC and NLRP3 contents were decreased in human NP cells after transfection with ASC siRNA and NLRP3 siRNA, as confirmed by immunoblotting. (D, E) Protein level of GSDMD-N in human NP cells was increased in the lactate group and decreased in the ASC siRNA and NLRP3 groups; levels were detected by immunoblotting. (F) LDH release by human NP cells was detected by flow cytometry. (G, I) The pyroptosis level of human NP cells was increased in the lactate group and decreased in the ASC siRNA and NLRP3 siRNA groups, as detected by immunofluorescence staining. Scale bar 50 = µm. (H, J) The pyroptosis level of human NP cells was increased in the lactate group and decreased in the ASC siRNA and NLRP3 siRNA groups, as detected by flow cytometry. Data are represented as mean ± SD (n = 3). Significant differences between groups are indicated as *P < .01, compared with control group; # P < .01, compared with lactate group
Fig 2: Pyroptosis is activated in the lung tissues of mice infected with A. fumigatus after irradiation. a Representative western blot images showing changes in the levels of caspase 1-p20, gasdermin D (GSDMD-N), interleukin 18 (IL-18), IL-1ß cleaved, b–e and their statistical results. f–i mRNA expression of ASC, NLRP3, NLRC4, and AIM2. j, k Secretion of IL-18 and IL-1ß in BALF. Data are shown as the mean ± standard deviation (n = 8). *P < 0.05, **P < 0.01 versus the control group
Fig 3: Oxymatrine decreased the levels of NLRP3, Gsdmd-N, caspase-1, ASC, IL-1ß, and IL-18 in INS-1 cells under high glucose and high fat conditions.(A) Representative Western blotting bands of NLRP3, Gsdmd-N, caspase-1, and IL-1ß proteins were showed. (B) Analysis of the amount of NLRP3 proteins. (C) Analysis of the amount of caspase-1 proteins. (D) Analysis of the amount of IL-1ß proteins. (E) Analysis of the amount of Gsdmd-N proteins. (F) Representative western blotting bands of ASC proteins were showed; Oxymatrine: 10 µM. (G) Analysis of the amount of ASC proteins; Oxymatrine: 10 µM. (H) Analysis of the secretion of IL-1ß and IL-18; Oxymatrine: 10 µM. (I) Analysis of the activity of caspase-1; Oxymatrine: 10 µM. control: no treatment; HG: high glucose (30 mM glucose); PA: palmitic acid sodium (400 µM). Data are presented as mean ± SD and represent an average of three experiments. NLRP3, Nod-like receptor family pyrin domain containing 3; Gsdmd, Gasdermin D; caspase-1, cysteine aspartic acid-specific proteinase-1; ASC, apoptosis-associated speck-like protein containing a CARD; IL, interleukin. Data from Western blotting are normalized to control. **p < 0.01 vs. control, #p < 0.05, ##p < 0.01 vs. HG + PA; the experiments were repeated three times.
Fig 4: The inhibition of TLR4/MyD88/NF-?B/NLRP3 cascade by Tanshinone IIA.(A) The mRNA levels of TLR4 and NLRP3 in the groups (n = 8 for each group). (B) The protein levels of TLR4, Myd88, p-NF-?B p65, NLRP3, and ASC (n = 3 for each group). (C) Representative immunohistochemical pictures of TLR4 and NLRP3 in cardiac tissues. Scale bar = 50 µm. The obtained data was revealed as the mean ± SD. CME, coronary microembolization; Tan-L, CME + low-dose Tan IIA (10 mg/kg); Tan-H, CME + high-dose Tan IIA (20 mg/kg). ***p < 0.001 vs. the sham group; ###p < 0.001 vs. the CME group.
Fig 5: Association between the expression levels of NLRP3 inflammasome components and hepatitis B virus (HBV) DNA load. Analysis by immunohistochemistry of expression levels of NLRP3, ASC, and IL-1ß in tumor-adjacent tissues of HCC patients with or without HBV infection. Representative images are shown, and the mean integrated optical density (IOD) of protein expression was statistically analyzed, n = 51 (a–d). Pearson correlation analysis of NLRP3, ASC, and IL-1ß expression and HBV DNA copy number in a cohort of 51 patients, n = 51 (e–g). Serum ASC in patients with varied HBV DNA load was detected by ELISA, n = 84 (h). The height of the histogram represents the DNA copy number counts. Data are shown as mean ± SD. *P < 0.05 and **P < 0.01
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