Journal: Nature immunology

Article Title: Anaphylactic degranulation by mast cells requires the mobilization of inflammasome components.

doi: 10.1038/s41590-024-01788-y

Figure Lengend Snippet: Fig. 1 | Inflammasome components mediate MC degranulation following IgE–Ag stimulation. a, Body temperature changes in wild-type (WT), Asc−/− and Nlrp3−/− mice and in KitW-sh/W-sh mice repleted with BMMCs from WT, Asc−/− or Nlrp3−/− mice sensitized with TNP-specific IgE antibody and injected i.v. with TNP-OVA and monitored for 105 min. b, β-hexosaminidase release from DNP- specific IgE-sensitized WT, Asc−/−, Nlrp3−/− and Casp-1/Casp-11−/− BMMCs after exposure to increasing doses of BSA-DNP. c, β-hexosaminidase release from IgE- sensitized and BSA-DNP stimulated RBL-2H3 MCs in which the expression of Asc, Nlrp3 or both was silenced with siRNA and immunoblot analysis of NLRP3 and ASC expression in each condition, with GAPDH as a loading control. d, β-hexosaminidase release from IgE-sensitized, BSA-DNP-stimulated Asc−/− or Nlrp3−/− BMMCs transduced with plasmids encoding mouse ASC, NLRP3 or empty vector (EV) and an immunoblot of endogenous NLRP3 and ASC and Flag–NLRP3 and Flag–ASC in cell lysates of each experimental condition. e, Measurement of intracellular Ca2+ concentrations using Fluo-4 NW assay kit in WT, Asc−/− and Nlrp3−/− BMMCs after IgE–Ag or thapsigargin exposure. f, β-hexosaminidase release from WT, Asc−/− and Nlrp3−/− BMMCs following exposure to thapsigargin

Article Snippet: Ectopic expression of NLRP3 and ASC in Nlrp3−/− or Asc−/− BMMCs BMMCs obtained from either Nlrp3−/− or Asc−/− (2 × 107) or WT were nucleofected with 5 μg of each expression plasmid expressing mouse Flag-tagged NLRP3 (Addgene, 75127) or ASC (Addgene, 75134) or empty plasmid using P3 Primary Cell 4D-Nucleofector X-kit (Lonza).

Techniques: Injection, Expressing, Western Blot, Control, Transduction, Plasmid Preparation

Journal: Nature immunology

Article Title: Anaphylactic degranulation by mast cells requires the mobilization of inflammasome components.

doi: 10.1038/s41590-024-01788-y

Figure Lengend Snippet: Fig. 2 | NLRP3 and ASC form a distinct complex with CD63 on MC granules. a, Immunofluorescence staining of WT BMMCs with CD63 (green), NLRP3 (red, left) or CD63 (green) and ASC (orange, right) at 0, 5 and 10 min following IgE–Ag stimulation. Scale bars, 20 μm. b, Immunoblotting of immunoprecipitated endogenous CD63 with endogenous NLRP3, ASC and caspase-1 in the immunoprecipitation (IP) fractions from untreated or IgE–Ag stimulated WT BMMCs. IgG antibody from the corresponding rabbit or mouse species was used as a control. Protein of interests in the total cell lysates (TCL) are depicted in each fraction. c, Immunoblot analyses of NLRP3, ASC, caspase-1 and CD63 in cell fractions from untreated, IgE–Ag-stimulated or LPS/nigericin-stimulated

Article Snippet: Ectopic expression of NLRP3 and ASC in Nlrp3−/− or Asc−/− BMMCs BMMCs obtained from either Nlrp3−/− or Asc−/− (2 × 107) or WT were nucleofected with 5 μg of each expression plasmid expressing mouse Flag-tagged NLRP3 (Addgene, 75127) or ASC (Addgene, 75134) or empty plasmid using P3 Primary Cell 4D-Nucleofector X-kit (Lonza).

Techniques: Immunofluorescence, Staining, Western Blot, Immunoprecipitation, Control

Journal: Nature immunology

Article Title: Anaphylactic degranulation by mast cells requires the mobilization of inflammasome components.

doi: 10.1038/s41590-024-01788-y

Figure Lengend Snippet: Fig. 3 | NEK7 and Pyk2 kinases are critical initiators of granulosome formation. a, Immunoblotting of NEK7 with NLRP3 from WT BMMCs stimulated with IgE–Ag or left untreated. Protein of interests in the TCL are depicted in each fraction. b, Immunoblot of NEK7 with NLRP3, ASC and CD63 in non-stimulated or IgE–Ag-stimulated WT BMMCs pretreated or not with CAY-10736 or oridonin. Protein of interests in the TCL are depicted in each fraction. c, Immunoblot analysis of cross-linked NLRP3 from IgE–Ag-stimulated or non-stimulated WT BMMCs pretreated or not with CAY-10736 or oridonin. d, Immunoblot analysis of CD63 interaction with NEK7, NLRP3 and ASC in unstimulated or IgE–Ag-stimulated WT BMMCs pretreated or not with CAY-10736 or oridonin. The proteins of interest in the TCL are depicted in each fraction. e, Secretion of β-hexosaminidase in WT and Nlrp3−/− BMMCs pretreated with vehicle, CAY- 10736 or oridonin and stimulated with IgE–Ag. f, Immunoblot analysis of PyK2 interaction with ASC in WT BMMCs at 0, 3, 5, 15 and 30 min after stimulation with IgE–Ag. The proteins of interest in the TCL are depicted in each fraction. g, Immunoblot analysis of PyK2 interactions with ASC, CD63 and NLRP3 in

Article Snippet: Ectopic expression of NLRP3 and ASC in Nlrp3−/− or Asc−/− BMMCs BMMCs obtained from either Nlrp3−/− or Asc−/− (2 × 107) or WT were nucleofected with 5 μg of each expression plasmid expressing mouse Flag-tagged NLRP3 (Addgene, 75127) or ASC (Addgene, 75134) or empty plasmid using P3 Primary Cell 4D-Nucleofector X-kit (Lonza).

Techniques: Western Blot

Journal: Nature immunology

Article Title: Anaphylactic degranulation by mast cells requires the mobilization of inflammasome components.

doi: 10.1038/s41590-024-01788-y

Figure Lengend Snippet: Fig. 4 | Granulosomes facilitate MT polymerization and granule trafficking. a, Immunofluorescence microscopy of α-tubulin staining in WT, Nlrp3−/− and Asc−/− BMMCs before and 10 min after IgE–Ag stimulation. Scale bars, 20 μm. b, Protein immunoblot analysis of insoluble and Triton buffer soluble fractions in WT, Nlrp3−/−, Asc−/− and Casp-1/11−/− BMMCs stimulated or not with IgE–Ag with SDS–PAGE gel stained with Coomassie brilliant blue used as an equal loading control. c, Immunoblot analysis of NLRP3, ASC or CD63 interactions with dynein in WT BMMCs stimulated with IgE–Ag for 0, 5 or 10 min. The proteins in the TCL are depicted in each fraction. d, Immunoblot analysis of CD63 interactions with

Article Snippet: Ectopic expression of NLRP3 and ASC in Nlrp3−/− or Asc−/− BMMCs BMMCs obtained from either Nlrp3−/− or Asc−/− (2 × 107) or WT were nucleofected with 5 μg of each expression plasmid expressing mouse Flag-tagged NLRP3 (Addgene, 75127) or ASC (Addgene, 75134) or empty plasmid using P3 Primary Cell 4D-Nucleofector X-kit (Lonza).

Techniques: Immunofluorescence, Microscopy, Staining, Western Blot, SDS Page, Control

Journal: Nature immunology

Article Title: Anaphylactic degranulation by mast cells requires the mobilization of inflammasome components.

doi: 10.1038/s41590-024-01788-y

Figure Lengend Snippet: Fig. 5 | NLRP3-targeting drug protects against anaphylaxis. a, β-hexosaminidase release in WT, Nlrp3−/− and Asc−/− BMMCs pretreated with 0, 2,10 and 50 μM of CY-09 and stimulated with IgE–Ag (TNP-OVA). b, Body temperature changes at 0, 30, 60, 90 and 120 min after Ag administration in IgE- sensitized KitW-sh/W-sh mice repleted with WT, Asc−/− or Nlrp3−/− BMMCs pretreated or not with CY-09 for 1 h. c, Immunoblot analysis of CD63 interactions with NLRP3, ASC and tubulin in non-stimulated or IgE–Ag-stimulated WT BMMCs pretreated or not with CY-09. The proteins of interest in TCL are depicted in each fraction. d, β-hexosaminidase release from Nlrp3 KD or Asc KD human LAD2 MCs pretreated or not with CY-09 and stimulated or not with IgE–Ag and immunoblots showing NLRP3 and ASC protein expression in TCL. Data

Article Snippet: Ectopic expression of NLRP3 and ASC in Nlrp3−/− or Asc−/− BMMCs BMMCs obtained from either Nlrp3−/− or Asc−/− (2 × 107) or WT were nucleofected with 5 μg of each expression plasmid expressing mouse Flag-tagged NLRP3 (Addgene, 75127) or ASC (Addgene, 75134) or empty plasmid using P3 Primary Cell 4D-Nucleofector X-kit (Lonza).

Techniques: Western Blot, Expressing

Journal: Nature immunology

Article Title: Anaphylactic degranulation by mast cells requires the mobilization of inflammasome components.

doi: 10.1038/s41590-024-01788-y

Figure Lengend Snippet: Fig. 6 | Pro-IL-1β is released from intact granules following IgE–Ag activation of LPS-primed MCs. a, ELISA showing release of IL-1β or β-hexosaminidase from WT or Nlrp3−/− BMMCs pretreated or not with TNP-specific IgE and LPS followed by stimulation with OVA-TNP (Ag) or not. b, Immunofluorescence staining of IL-1β (red) and CD63 (green) in WT or Nlrp3−/− BMMCs primed with LPS and TNP-specific IgE for 3–4 h and stimulated with OVA-TNP (Ag) at 0, 5 and 10 min (WT) or 10 min (Nlrp3−/−) and colocalization of IL-1β with CD63 calculated from 13 representative images using ImageJ (Pearson correlation coefficient). Values are the mean ± s.e.m.; significant differences of treatment versus control were analyzed by one-way ANOVA/Bonferroni’s post hoc test, ***P < 0.001, n = 13 images per group. Scale bars, 20 μm. c, Immunoblot analysis of pro-IL-1β and mature IL-1β in concentrated supernatants of WT or Nlrp3−/− BMMCs activated with IgE–Ag alone or LPS + IgE–Ag and Nlrp3−/− BMMCs transfected with Flag or Flag-tagged mouse NLRP3. d, Immunoblot analysis of pro-IL-1β and mature IL-1β in the supernatant of non-primed or LPS + IgE-primed Ag-activated WT BMMCs pretreated or not with chymostatin. e, Immunoblot analysis of pro-IL-1β and mature IL-1β in the supernatant and granule remnant fractions of WT BMMCs primed or not with LPS and stimulated or not with Ag. f, Body temperature changes at 0, 30, 60, 90 and 120 min after Ag administration in WT mice pre- sensitized with TNP-specific IgE antibody (5 µg per mouse) for 2–4 days before

Article Snippet: Ectopic expression of NLRP3 and ASC in Nlrp3−/− or Asc−/− BMMCs BMMCs obtained from either Nlrp3−/− or Asc−/− (2 × 107) or WT were nucleofected with 5 μg of each expression plasmid expressing mouse Flag-tagged NLRP3 (Addgene, 75127) or ASC (Addgene, 75134) or empty plasmid using P3 Primary Cell 4D-Nucleofector X-kit (Lonza).

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Control, Western Blot, Transfection

Journal: Journal of Cellular and Molecular Medicine

Article Title: Anti‐inflammatory effect of Trichospira verticillata via suppression of the NLRP3 inflammasome in neutrophilic asthma

doi: 10.1111/jcmm.18356

Figure Lengend Snippet: TVE specifically suppresses NLRP3 inflammasome. (A) Schematic outline of the NLRP3 inflammasome activation experiments in vitro. (B–C) PMA (500 nM)‐differentiated THP‐1 cells (B) and J774A.1 cells (C) were primed by LPS (100 ng/mL) for 3 h, before were treated with TVE for 2 h. Cell viability was analysed by EZ‐Cytox. (D–E) LPS‐primed THP‐1 cells were treated with TVE for 2 h and then stimulated for 1 h with nigericin (10 μM) (D) and ATP (5 mM) (E) with or without MCC950. (F–G) LPS‐primed J774A.1 cells were treated with TVE for 2 h and then stimulated for 30 min with nigericin (10 μM) (F), ATP (5 mM) (G) with or without MCC950. This data represents the investigation of the protein expression level of IL‐1β in supernatant, confirmed by immunoblot, using densitometry to assess intensity. (H‐J) IL‐1β (p17) and in the supernatants (Sup) and soluble lysates (Lys) were analysed by immunoblot. LPS‐primed J774A.1 cells were treated with TVE for 2 h and then stimulated for 3 h with MSU (100 μg/mL) (H) or dsDNA (2 μg/mL) (I) and flagellin (1.25 μg/mL) (J) by using Lipofectamine 3000™. One representative result of three independent experiments is shown. Values shown are reported as the means of technical triplicates ± SEM. One‐way ANOVA, Bonferroni post‐hoc test; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ATP, Adenosine triphosphate; IL‐1β, Interleukin‐1beta; LPS, Lipopolysaccharide; MCC, MCC950; Nig, Nigericin; n.s., not significant; TVE, Trichospira verticillata (L.) S.F. Blake Extract.

Article Snippet: The purified mouse NLRP3 (BPS bioscience, 100189) (0.139 mg/mL) was incubated with TVE (50 μg/mL) or DMSO at 37°C for 30 min in the reaction buffer (100 mM Tris pH 7.8, 2.8 mM EDTA, 100 mM MgCl₂, 15 mM KCl, 655 mM NaCl).

Techniques: Activation Assay, In Vitro, Expressing, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Anti‐inflammatory effect of Trichospira verticillata via suppression of the NLRP3 inflammasome in neutrophilic asthma

doi: 10.1111/jcmm.18356

Figure Lengend Snippet: TVE suppresses NLRP3 inflammasome activation regardless of K + efflux, ROS and ATPase activity. (A) LPS‐primed J774A.1 cells were treated with TVE for 2 h and then stimulated with IMQ (200 μM) for 1 h. (B) LPS‐primed J774A.1 cells were treated with or without TVE or NAC for 2 h and then stimulated with ATP (5 mM) for 25 min. ROS levels are detected by a microplate reader using a DCFDA solution (20 μM). (C) The amount of NLRP3‐mediated ATP converted into ADP with TVE was determined by luminescence using the ADP‐Glo Assay. (D) and (E) The NLRP3‐Myc transfected‐HEK 293FT cells were treated with or without TVE for 2 h. The NLRP3‐NEK7 interaction was analysed by immunoprecipitation and immunoblot. One representative result of three independent experiments is shown. Values are shown reported as the means of technical triplicates ± SEM. One‐way ANOVA, Bonferroni post‐hoc test; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. LPS, Lipopolysaccharide; n.s., not significant; NAC, N‐acetyl‐l‐cysteine; ROS, reactive oxygen species; TVE, Trichospira verticillata (L.) S.F. Blake Extract.

Article Snippet: The purified mouse NLRP3 (BPS bioscience, 100189) (0.139 mg/mL) was incubated with TVE (50 μg/mL) or DMSO at 37°C for 30 min in the reaction buffer (100 mM Tris pH 7.8, 2.8 mM EDTA, 100 mM MgCl₂, 15 mM KCl, 655 mM NaCl).

Techniques: Activation Assay, Activity Assay, Glo Assay, Transfection, Immunoprecipitation, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Anti‐inflammatory effect of Trichospira verticillata via suppression of the NLRP3 inflammasome in neutrophilic asthma

doi: 10.1111/jcmm.18356

Figure Lengend Snippet: TVE blocks NLRP3 inflammasome assembly by hindering ASC oligomerization. (A) One representative ASC speck (indicated by arrow) images of at least 10 images are shown. LPS‐primed J774A.1 cells were treated with TVE for 2 h and then stimulated with ATP (5 mM) for 30 min. (B) LPS‐primed THP‐1 cells were treated with TVE for 2 h and then stimulated with nigericin (10 μM) for 30 min. ASC oligomerization in the cross‐linked cytosolic pellet was analysed by immunoblot. Scale bar, 20 μm. One representative result of three independent experiments is shown. Values are shown reported as the means of 10 replicates ± SEM. One‐way ANOVA, Bonferroni post‐hoc test; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ASC, apoptosis‐associated speck‐like protein; LPS, Lipopolysaccharide; n.s., not significant; TVE, Trichospira verticillata (L.) S.F. Blake Extract.

Article Snippet: The purified mouse NLRP3 (BPS bioscience, 100189) (0.139 mg/mL) was incubated with TVE (50 μg/mL) or DMSO at 37°C for 30 min in the reaction buffer (100 mM Tris pH 7.8, 2.8 mM EDTA, 100 mM MgCl₂, 15 mM KCl, 655 mM NaCl).

Techniques: Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Anti‐inflammatory effect of Trichospira verticillata via suppression of the NLRP3 inflammasome in neutrophilic asthma

doi: 10.1111/jcmm.18356

Figure Lengend Snippet: TVE inhibits NLRP3 inflammasome in lung epithelial cells. (A–B) A549 cells were treated with TVE for 2 h (A), or overnight (B). Cell viability was analysed by EZ‐CYTOX. (C–D) A549 cells and PMA (500 nM)‐differentiated THP‐1 cells were co‐cultured and primed by LPS (100 ng/mL) for 3 h, before they were treated with TVE for 2 h and stimulated for 30 min with nigericin (10 μM) (C), and ATP (5 mM) (D). The level of IL‐1β was evaluated by ELISA and the lysate was analysed by immunoblotting. One representative result of three independent experiments is shown. Values are shown reported as the means of technical triplicates ± SEM. One‐way ANOVA, Bonferroni post‐hoc test; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n.s. not significant.

Article Snippet: The purified mouse NLRP3 (BPS bioscience, 100189) (0.139 mg/mL) was incubated with TVE (50 μg/mL) or DMSO at 37°C for 30 min in the reaction buffer (100 mM Tris pH 7.8, 2.8 mM EDTA, 100 mM MgCl₂, 15 mM KCl, 655 mM NaCl).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Nature

Article Title: DDX3X acts as a live-or-die checkpoint in stressed cells by regulating NLRP3 inflammasome

doi: 10.1038/s41586-019-1551-2

Figure Lengend Snippet: a, Co-immunoprecipitation (IP) of NLRP3 and DDX3X in BMDMs treated with LPS with or without nigericin. Representative blots (n= 3). b, Immunoblot analysis of CASP1 cleavage in wild-type (WT), Ddx3xfl/fl, Ddx3xfl/flLysMcre and Nlrp3−/− BMDMs treated with LPS and nigericin. Representative blots (n > 3). c, Immunoblots of DDX3X expression and CASP1 cleavage after siRNA-mediated knockdown of Ddx3x in BMDMs treated with LPS with or without nigericin. Representative blots (n = 2). d, Confocal microscopy imaging of ASC specks in BMDMs treated with LPS with or without nigericin, to visualize the subcellular localization of DDX3X, NLRP3 and ASC. Scale bars, 10 μm. Representative images (n = 3). e, Schematic of N-terminal Flag-tagged NLRP3 expression constructs: full-length NLRP3 (Flag-NLRP3-FL), the pyrin domain of NLRP3 (Flag-PYD; amino acids 1–90 of NLRP3), the NACHT domain of NLRP3 (Flag-NACHT; amino acids 91–710 of NLRP3) and the LRR domain of NLRP3 (Flag-LRR; amino acids 711–1034 of NLRP3). f, Immunoblot (IB) analysis of input lysates used for co-immunoprecipitation of Flag-tagged NLRP3 constructs and DDX3X-mCherry. Representative blots (n > 3). g, Immunoblot analysis of immunoprecipitation of DDX3X-mCherry and the indicated NLRP3 constructs. Red asterisk indicates the antibody light chain. Representative blots (n > 3).

Article Snippet: NLRP3 expression constructs pCDNA3-N-Flag-NLRP3, pCDNA3-N-Flag-NLRP3 1–90, pCDNA3-N-Flag-NLRP3 91–710 and pCDNA3-N-Flag-NLRP3 711–1034 were purchased from AddGene.

Techniques: Immunoprecipitation, Western Blot, Expressing, Confocal Microscopy, Imaging, Construct

Journal: Nature

Article Title: DDX3X acts as a live-or-die checkpoint in stressed cells by regulating NLRP3 inflammasome

doi: 10.1038/s41586-019-1551-2

Figure Lengend Snippet: a, Immunoblot analysis of lysates fractionated by analytical ultra-centrifugation to test the oligomerization of NLRP3, DDX3X and ASC in BMDMs stimulated with LPS and treated either with nigericin or with arsenite and nigericin. Representative blots (n = 2). MW, molecular weight. b, Immunoblot analysis of ASC from the insoluble fraction of samples from a under non-reducing (without β-mercaptoethanol; – BME) and reducing (+BME) conditions. Representative blots (n = 2). c, STORM imaging of ASC speck assembly and its spatial organization with DDX3X over time. Scale bars, 5 μm (whole-cell images); 1 μm (magnified images). Representative images (n = 2). d, Plot of the r-index to show the extent of colocalization of ASC and DDX3X with the duration of nigericin treatment (n = 2). R2 was calculated using linear regression analysis; P = 0.0125 (for the significance of the slope being non-zero). Data are mean ± s.e.m. e, f, Immunoblot analysis of CASP1 cleavage in LPS-primed BMDMs to which arsenite was added at various time points (30 and 15 min before adding nigericin, simultaneously with nigericin and 15 and 30 min after adding nigericin), without (e) or with (f) pre-treatment with anisomycin. Representative blots (n = 3). g, h, Structured illumination microscopy of BMDMs to visualize the subcellular localization of DDX3X, G3BP1 and ASC in LPS-primed BMDMs treated with nigericin alone (g) or with arsenite and nigericin (h). Scale bars, 3 μm. Representative images (n = 3).

Article Snippet: NLRP3 expression constructs pCDNA3-N-Flag-NLRP3, pCDNA3-N-Flag-NLRP3 1–90, pCDNA3-N-Flag-NLRP3 91–710 and pCDNA3-N-Flag-NLRP3 711–1034 were purchased from AddGene.

Techniques: Western Blot, Centrifugation, Molecular Weight, Imaging, Microscopy

Journal: Nature

Article Title: DDX3X acts as a live-or-die checkpoint in stressed cells by regulating NLRP3 inflammasome

doi: 10.1038/s41586-019-1551-2

Figure Lengend Snippet: a, Confocal microscopy imaging of peritoneal CD45+ cells to visualize in vivo stress granules in Ddx3xfl/fl and Ddx3xfl/flLysMcre mice that were treated with PBS or arsenite. Scale bars, 10 μm. Representative images (n = 2). b, Quantification of CD45+ cells that contain stress granules, from the peritoneal cavity of Ddx3xfl/fl and Ddx3xfl/flLysMcre mice that were injected with arsenite or PBS. ****P < 0.0001 (unpaired two-sided t-test). Data represent two biologically independent experiments (n = 10 frames). c, Quantification of the numbers and percentage of CD11b+ myeloid cells in the peritoneal cavity. P values (from left to right): ***P = 0.0001, ***P = 0.0005 (unpaired two-sided t-test; n = 7). d, e, Levels of IL-1β in the serum (d) and peritoneal fluid (PF) (e) of mice that were injected with LPS with or without prior arsenite injection in the peritoneum. P values: **P = 0.0026 (d), P = 0.09 (e) (unpaired two-sided t-test; n = 7). f, Levels of IL-1β in the serum and peritoneal fluid of Ddx3xfl/fl and Ddx3xfl/flLysMcre mice that were injected with LPS in the peritoneum. P values (from top to bottom): **P = 0.0073, *P = 0.0123 (unpaired two-sided t-test; n ≥ 6). Data are mean ± s.e.m. (b-f). g, Schematic of the interplay between the inflammasome and stress granules that is involved in cell-fate decisions. DDX3X promotes NLRP3 inflammasome activation and the pro-death cell-fate decision probably by interacting with the NLRP3 NACHT domain through its helicase (Heli) domain. Induction of stress granules causes the sequestration of DDX3X (along with 40S ribosomal subunits and translation initiation factors (eIFs)), thus making it unavailable for NLRP3 inflammasome activation and thereby allowing the cells to make a pro-survival cell-fate choice.

Article Snippet: NLRP3 expression constructs pCDNA3-N-Flag-NLRP3, pCDNA3-N-Flag-NLRP3 1–90, pCDNA3-N-Flag-NLRP3 91–710 and pCDNA3-N-Flag-NLRP3 711–1034 were purchased from AddGene.

Techniques: Confocal Microscopy, Imaging, In Vivo, Injection, Activation Assay