Review



primary anti nlrp3  (Proteintech)


Bioz Verified Symbol Proteintech is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 96
      Buy from Supplier

    Structured Review

    Proteintech primary anti nlrp3
    Primary Anti Nlrp3, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1647 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nlrp3+primary+antibody/NLRP3+Antibody/pm41698560-70-4-13
    Average 96 stars, based on 1647 article reviews
    primary anti nlrp3 - by Bioz Stars, 2026-09
    96/100 stars

    Images

    Related Articles

    Incubation:

    Article Title: Size-dependent pyroptosis induction by calcium oxalate monohydrate crystals in HK-2 cells
    Article Snippet: .. After reaching the incubation time, the culture medium was removed, washed twice with PBS, fixed with paraformaldehyde for 15 min, blocked with sheep serum for 20 min, and then incubated with NLRP3 primary antibody (1:100) (Proteintech, Cat No. 30109-1-AP) at 4°C overnight. .. Upon arrival of the incubation time, the cells were washed three times with PBS, then incubated at 37°C for 0.5 h with FITC secondary antibody drop in the dark, and finally labeled with DAPI staining (Shanghai Beyotime Biotechnology, Cat No. C1006-10 mL) for 10min.



    Similar Products

    primary antibodies against nlrp3  (Affinity Biosciences)
    86
    Affinity Biosciences primary antibodies against nlrp3
    Primary Antibodies Against Nlrp3, supplied by Affinity Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nlrp3+primary+antibody/anti+nlrp3/pm42259094-94-22-57
    Average 86 stars, based on 1 article reviews
    primary antibodies against nlrp3 - by Bioz Stars, 2026-09
    86/100 stars
    		  Buy from Supplier
    primary antibodies against nlrp3  (Cell Signaling Technology Inc)
    98
    Cell Signaling Technology Inc primary antibodies against nlrp3
    Composite hydrogel promotes the polarization of BV2 cells to M2 types and alleviates PC12 cell pyroptosis in vitro inflammatory environment. (A) Representative western blots showing protein expression of iNOS and Arg-1 in each group, β-actin was utilized as a loading control. (B) Quantitative analysis of relative expression of iNOS and Arg-1. (C) Representative immunofluorescence images of CD68 positive and iNOS positive BV2 cells (scale bar: 20 μm). (D) Representative immunofluorescence images of CD68 positive and Arg-1 positive BV2 cells (scale bar: 20 μm). (E, F) Quantitative analysis of relative fluorescence intensity of iNOS and Arg-1. (G) Representative western blots showing the expression of <t>NLRP3,</t> Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein associated with pyroptosis, β-actin was utilized as a loading control. (H) PI staining of PC12 cells in each group (scale bar, 100 μm). (I) Quantitative analysis of PI staining of PC12 cells (n = 3). (J) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.
    Primary Antibodies Against Nlrp3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nlrp3+primary+antibody/NLRP3+Rabbit+mAb/pmc12907102-312-0-10
    Average 98 stars, based on 1 article reviews
    primary antibodies against nlrp3 - by Bioz Stars, 2026-09
    98/100 stars
    		  Buy from Supplier
    primary antibody nlrp3  (Abmart Inc)
    86
    Abmart Inc primary antibody nlrp3
    Composite hydrogel promotes the polarization of BV2 cells to M2 types and alleviates PC12 cell pyroptosis in vitro inflammatory environment. (A) Representative western blots showing protein expression of iNOS and Arg-1 in each group, β-actin was utilized as a loading control. (B) Quantitative analysis of relative expression of iNOS and Arg-1. (C) Representative immunofluorescence images of CD68 positive and iNOS positive BV2 cells (scale bar: 20 μm). (D) Representative immunofluorescence images of CD68 positive and Arg-1 positive BV2 cells (scale bar: 20 μm). (E, F) Quantitative analysis of relative fluorescence intensity of iNOS and Arg-1. (G) Representative western blots showing the expression of <t>NLRP3,</t> Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein associated with pyroptosis, β-actin was utilized as a loading control. (H) PI staining of PC12 cells in each group (scale bar, 100 μm). (I) Quantitative analysis of PI staining of PC12 cells (n = 3). (J) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.
    Primary Antibody Nlrp3, supplied by Abmart Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nlrp3+primary+antibody/anti+nlrp3/pm41903063-97-23-27
    Average 86 stars, based on 1 article reviews
    primary antibody nlrp3 - by Bioz Stars, 2026-09
    86/100 stars
    		  Buy from Supplier
    nlrp3 primary antibody  (Servicebio Inc)
    86
    Servicebio Inc nlrp3 primary antibody
    Composite hydrogel promotes the polarization of BV2 cells to M2 types and alleviates PC12 cell pyroptosis in vitro inflammatory environment. (A) Representative western blots showing protein expression of iNOS and Arg-1 in each group, β-actin was utilized as a loading control. (B) Quantitative analysis of relative expression of iNOS and Arg-1. (C) Representative immunofluorescence images of CD68 positive and iNOS positive BV2 cells (scale bar: 20 μm). (D) Representative immunofluorescence images of CD68 positive and Arg-1 positive BV2 cells (scale bar: 20 μm). (E, F) Quantitative analysis of relative fluorescence intensity of iNOS and Arg-1. (G) Representative western blots showing the expression of <t>NLRP3,</t> Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein associated with pyroptosis, β-actin was utilized as a loading control. (H) PI staining of PC12 cells in each group (scale bar, 100 μm). (I) Quantitative analysis of PI staining of PC12 cells (n = 3). (J) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.
    Nlrp3 Primary Antibody, supplied by Servicebio Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nlrp3+primary+antibody/anti+nlrp3/pm41754836-389-20-23
    Average 86 stars, based on 1 article reviews
    nlrp3 primary antibody - by Bioz Stars, 2026-09
    86/100 stars
    		  Buy from Supplier
    primary anti nlrp3  (Proteintech)
    96
    Proteintech primary anti nlrp3
    Composite hydrogel promotes the polarization of BV2 cells to M2 types and alleviates PC12 cell pyroptosis in vitro inflammatory environment. (A) Representative western blots showing protein expression of iNOS and Arg-1 in each group, β-actin was utilized as a loading control. (B) Quantitative analysis of relative expression of iNOS and Arg-1. (C) Representative immunofluorescence images of CD68 positive and iNOS positive BV2 cells (scale bar: 20 μm). (D) Representative immunofluorescence images of CD68 positive and Arg-1 positive BV2 cells (scale bar: 20 μm). (E, F) Quantitative analysis of relative fluorescence intensity of iNOS and Arg-1. (G) Representative western blots showing the expression of <t>NLRP3,</t> Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein associated with pyroptosis, β-actin was utilized as a loading control. (H) PI staining of PC12 cells in each group (scale bar, 100 μm). (I) Quantitative analysis of PI staining of PC12 cells (n = 3). (J) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.
    Primary Anti Nlrp3, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nlrp3+primary+antibody/NLRP3+Antibody/pm41698560-70-4-13
    Average 96 stars, based on 1 article reviews
    primary anti nlrp3 - by Bioz Stars, 2026-09
    96/100 stars
    		  Buy from Supplier
    primary antibodies for nlrp3  (Adipogen)
    86
    Adipogen primary antibodies for nlrp3
    Composite hydrogel promotes the polarization of BV2 cells to M2 types and alleviates PC12 cell pyroptosis in vitro inflammatory environment. (A) Representative western blots showing protein expression of iNOS and Arg-1 in each group, β-actin was utilized as a loading control. (B) Quantitative analysis of relative expression of iNOS and Arg-1. (C) Representative immunofluorescence images of CD68 positive and iNOS positive BV2 cells (scale bar: 20 μm). (D) Representative immunofluorescence images of CD68 positive and Arg-1 positive BV2 cells (scale bar: 20 μm). (E, F) Quantitative analysis of relative fluorescence intensity of iNOS and Arg-1. (G) Representative western blots showing the expression of <t>NLRP3,</t> Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein associated with pyroptosis, β-actin was utilized as a loading control. (H) PI staining of PC12 cells in each group (scale bar, 100 μm). (I) Quantitative analysis of PI staining of PC12 cells (n = 3). (J) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.
    Primary Antibodies For Nlrp3, supplied by Adipogen, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nlrp3+primary+antibody/anti+nlrp3/pm41664375-69-0-4
    Average 86 stars, based on 1 article reviews
    primary antibodies for nlrp3 - by Bioz Stars, 2026-09
    86/100 stars
    		  Buy from Supplier
    primary antibodies for nlrp3  (Cell Signaling Technology Inc)
    86
    Cell Signaling Technology Inc primary antibodies for nlrp3
    RBG Inhibits <t>NLRP3</t> Inflammasome Assembly and Inflammation in Atherosclerotic Plaques of ApoE -/- Mice. (A) Volcano plot of differentially expressed genes between the RBG-H group and HFD group (n = 3). (B) GO enrichment analysis of genes with differential expression. (C) KEGG enrichment analysis of genes with differential expression. (E, F, G, H) Serum levels of IL18, IL1β, TNF-α, and IL10 were measured (n = 10). (D, I, J, K, L, M, N, O, P) Western blot analysis and Statistical analysis was conducted for NLRP3, ASC total protein, ASC oligomerization, NEK7, Caspase-1, Cleaved Caspase-1, Pro IL18, and IL18 (n = 6). (Q, R, S, T, U) Representative images and statistical analysis of immunofluorescence staining for NLRP3, ASC, and Caspase-1 in the aortic sinus (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns indicates not significant.
    Primary Antibodies For Nlrp3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nlrp3+primary+antibody/pmc12869210-47-0-27
    Average 86 stars, based on 1 article reviews
    primary antibodies for nlrp3 - by Bioz Stars, 2026-09
    86/100 stars
    		  Buy from Supplier
    nlrp3 primary antibody  (Proteintech)
    96
    Proteintech nlrp3 primary antibody
    Differences in pyroptosis induced by COM crystals of different sizes (A) Caspase-1/PI double staining flow cytometry quantitative analysis. (B) Caspase-1/PI/Hoechst 33342 triple staining confocal observation. Scale bars, 50 μm. (C) <t>NLRP3</t> immunofluorescence map. Scale bars, 100 μm. (D) Quantitative histogram of pyroptosis; n = 3; mean ± SEM. (E) Quantitative bar graph of NLRP3; n = 3; mean ± SEM. NC: normal control. COM crystals were incubated with HK-2 cells for 48 h. Compared with NC group, ∗ p < 0.05; ∗∗ p < 0.01. (A) flow graph is divided into four regions (Q1, Q2, Q3, and Q4), of which Q1 is PI high signal area and Caspase-1 low signal area, representing apoptotic necrotic cells. Q2 is the PI high signal region and Caspase-1 high signal region, which represents the late pyroptosis cells. Q3 is the PI low signal area and Caspase-1 high signal area, which represents the early pyroptosis cells (Caspase-1 is actively expressed). Q4 is the PI hypointense region and Caspase-1 hypointense region, representing normal cells. Q1+ Q2 refers to dead cells, and Q2+ Q3 refers to pyroptosis cells. (B) shows the presence of four types of cells: the first type is the normal cell (indicated by the orange arrow), which corresponds to the Q4 region cells in (A); the second type was apoptotic or necrotic cells (indicated by white arrow), namely Q1 region cells. The third type was the late pyroptosis cells (yellow arrow), which were Q2 cells. The fourth type is the cells in the early stage of pyroptosis (purple arrow), which is the Q3 region cells. Orange arrows refer to normal cells, white arrows to apoptotic necrotic cells, yellow arrows to late pyroptosis cells, and purple arrows to early pyroptosis cells.
    Nlrp3 Primary Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nlrp3+primary+antibody/NLRP3+Antibody/pmc12803861-258-31-35
    Average 96 stars, based on 1 article reviews
    nlrp3 primary antibody - by Bioz Stars, 2026-09
    96/100 stars
    		  Buy from Supplier
    primary antibodies against nlrp3  (Wanleibio)
    86
    Wanleibio primary antibodies against nlrp3
    Differences in pyroptosis induced by COM crystals of different sizes (A) Caspase-1/PI double staining flow cytometry quantitative analysis. (B) Caspase-1/PI/Hoechst 33342 triple staining confocal observation. Scale bars, 50 μm. (C) <t>NLRP3</t> immunofluorescence map. Scale bars, 100 μm. (D) Quantitative histogram of pyroptosis; n = 3; mean ± SEM. (E) Quantitative bar graph of NLRP3; n = 3; mean ± SEM. NC: normal control. COM crystals were incubated with HK-2 cells for 48 h. Compared with NC group, ∗ p < 0.05; ∗∗ p < 0.01. (A) flow graph is divided into four regions (Q1, Q2, Q3, and Q4), of which Q1 is PI high signal area and Caspase-1 low signal area, representing apoptotic necrotic cells. Q2 is the PI high signal region and Caspase-1 high signal region, which represents the late pyroptosis cells. Q3 is the PI low signal area and Caspase-1 high signal area, which represents the early pyroptosis cells (Caspase-1 is actively expressed). Q4 is the PI hypointense region and Caspase-1 hypointense region, representing normal cells. Q1+ Q2 refers to dead cells, and Q2+ Q3 refers to pyroptosis cells. (B) shows the presence of four types of cells: the first type is the normal cell (indicated by the orange arrow), which corresponds to the Q4 region cells in (A); the second type was apoptotic or necrotic cells (indicated by white arrow), namely Q1 region cells. The third type was the late pyroptosis cells (yellow arrow), which were Q2 cells. The fourth type is the cells in the early stage of pyroptosis (purple arrow), which is the Q3 region cells. Orange arrows refer to normal cells, white arrows to apoptotic necrotic cells, yellow arrows to late pyroptosis cells, and purple arrows to early pyroptosis cells.
    Primary Antibodies Against Nlrp3, supplied by Wanleibio, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nlrp3+primary+antibody/anti+nlrp3/pm41539751-38-0-11
    Average 86 stars, based on 1 article reviews
    primary antibodies against nlrp3 - by Bioz Stars, 2026-09
    86/100 stars
    		  Buy from Supplier

    Image Search Results


    Composite hydrogel promotes the polarization of BV2 cells to M2 types and alleviates PC12 cell pyroptosis in vitro inflammatory environment. (A) Representative western blots showing protein expression of iNOS and Arg-1 in each group, β-actin was utilized as a loading control. (B) Quantitative analysis of relative expression of iNOS and Arg-1. (C) Representative immunofluorescence images of CD68 positive and iNOS positive BV2 cells (scale bar: 20 μm). (D) Representative immunofluorescence images of CD68 positive and Arg-1 positive BV2 cells (scale bar: 20 μm). (E, F) Quantitative analysis of relative fluorescence intensity of iNOS and Arg-1. (G) Representative western blots showing the expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein associated with pyroptosis, β-actin was utilized as a loading control. (H) PI staining of PC12 cells in each group (scale bar, 100 μm). (I) Quantitative analysis of PI staining of PC12 cells (n = 3). (J) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.

    Journal: Bioactive Materials

    Article Title: 3D-MSCs apoptotic bodies-integrated conductive hydrogel mitigates spinal cord injury via immunoregulation and alleviating neuronal pyroptosis

    doi: 10.1016/j.bioactmat.2026.01.043

    Figure Lengend Snippet: Composite hydrogel promotes the polarization of BV2 cells to M2 types and alleviates PC12 cell pyroptosis in vitro inflammatory environment. (A) Representative western blots showing protein expression of iNOS and Arg-1 in each group, β-actin was utilized as a loading control. (B) Quantitative analysis of relative expression of iNOS and Arg-1. (C) Representative immunofluorescence images of CD68 positive and iNOS positive BV2 cells (scale bar: 20 μm). (D) Representative immunofluorescence images of CD68 positive and Arg-1 positive BV2 cells (scale bar: 20 μm). (E, F) Quantitative analysis of relative fluorescence intensity of iNOS and Arg-1. (G) Representative western blots showing the expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein associated with pyroptosis, β-actin was utilized as a loading control. (H) PI staining of PC12 cells in each group (scale bar, 100 μm). (I) Quantitative analysis of PI staining of PC12 cells (n = 3). (J) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.

    Article Snippet: Primary antibodies against NLRP3 (cat. no. 15101) were obtained from Cell Signaling Technology (Beverly, Massachusetts, USA).

    Techniques: In Vitro, Western Blot, Expressing, Control, Immunofluorescence, Fluorescence, Staining, Comparison

    The composite hydrogel inhibits post-SCI pyroptosis in neurons. (A) Immunofluorescence images of residual neurons existing in the anterior horn of the spinal cord (scale bar, 500 μm and 200 μm). (B) Immunofluorescence image of Caspase-1 expression of neurons 3 days after SCI (scale bar, 20 μm). (C) Quantitative analysis of relative fluorescence intensity of Caspase-1 protein expression in neurons within the specified groups (n = 3). (D) Immunofluorescence image of GSDMD-N expression of neurons 3 days after SCI (scale bar, 20 μm). (E) Quantitative analysis of relative fluorescence intensity of GSDMD-N protein expression in neurons within the specified groups (n = 3). (F) Representative western blots showing the expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein 3 days after SCI, GAPDH was utilized as a loading control. (G) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.

    Journal: Bioactive Materials

    Article Title: 3D-MSCs apoptotic bodies-integrated conductive hydrogel mitigates spinal cord injury via immunoregulation and alleviating neuronal pyroptosis

    doi: 10.1016/j.bioactmat.2026.01.043

    Figure Lengend Snippet: The composite hydrogel inhibits post-SCI pyroptosis in neurons. (A) Immunofluorescence images of residual neurons existing in the anterior horn of the spinal cord (scale bar, 500 μm and 200 μm). (B) Immunofluorescence image of Caspase-1 expression of neurons 3 days after SCI (scale bar, 20 μm). (C) Quantitative analysis of relative fluorescence intensity of Caspase-1 protein expression in neurons within the specified groups (n = 3). (D) Immunofluorescence image of GSDMD-N expression of neurons 3 days after SCI (scale bar, 20 μm). (E) Quantitative analysis of relative fluorescence intensity of GSDMD-N protein expression in neurons within the specified groups (n = 3). (F) Representative western blots showing the expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein 3 days after SCI, GAPDH was utilized as a loading control. (G) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.

    Article Snippet: Primary antibodies against NLRP3 (cat. no. 15101) were obtained from Cell Signaling Technology (Beverly, Massachusetts, USA).

    Techniques: Immunofluorescence, Expressing, Fluorescence, Western Blot, Control, Comparison

    RBG Inhibits NLRP3 Inflammasome Assembly and Inflammation in Atherosclerotic Plaques of ApoE -/- Mice. (A) Volcano plot of differentially expressed genes between the RBG-H group and HFD group (n = 3). (B) GO enrichment analysis of genes with differential expression. (C) KEGG enrichment analysis of genes with differential expression. (E, F, G, H) Serum levels of IL18, IL1β, TNF-α, and IL10 were measured (n = 10). (D, I, J, K, L, M, N, O, P) Western blot analysis and Statistical analysis was conducted for NLRP3, ASC total protein, ASC oligomerization, NEK7, Caspase-1, Cleaved Caspase-1, Pro IL18, and IL18 (n = 6). (Q, R, S, T, U) Representative images and statistical analysis of immunofluorescence staining for NLRP3, ASC, and Caspase-1 in the aortic sinus (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns indicates not significant.

    Journal: Journal of Advanced Research

    Article Title: Resibufogenin protects against atherosclerosis in ApoE -/- mice through blocking NLRP3 inflammasome assembly

    doi: 10.1016/j.jare.2025.04.029

    Figure Lengend Snippet: RBG Inhibits NLRP3 Inflammasome Assembly and Inflammation in Atherosclerotic Plaques of ApoE -/- Mice. (A) Volcano plot of differentially expressed genes between the RBG-H group and HFD group (n = 3). (B) GO enrichment analysis of genes with differential expression. (C) KEGG enrichment analysis of genes with differential expression. (E, F, G, H) Serum levels of IL18, IL1β, TNF-α, and IL10 were measured (n = 10). (D, I, J, K, L, M, N, O, P) Western blot analysis and Statistical analysis was conducted for NLRP3, ASC total protein, ASC oligomerization, NEK7, Caspase-1, Cleaved Caspase-1, Pro IL18, and IL18 (n = 6). (Q, R, S, T, U) Representative images and statistical analysis of immunofluorescence staining for NLRP3, ASC, and Caspase-1 in the aortic sinus (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns indicates not significant.

    Article Snippet: Primary antibodies for NLRP3 (ab15101), NF-κB (ab4764), NEK7 (ab10054), ASC (ab67824), Caspase-1 (ab24232), IL18 (ab57058), IL1β (ab31202), CD68 (ab97778), GAPDH (ab2118), and β-ACTIN (ab4970) were obtained from Cell Signaling Technology, Denvers, MA, USA.The antibodies PE Anti-Mouse CD86 (GL-1), APC Anti-Mouse CD206/MMR (C068C2), and PE/Cyanine7 Anti-Mouse F4/80 (CI A3-1) were procured from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

    Techniques: Quantitative Proteomics, Western Blot, Immunofluorescence, Staining

    RBG enhances foam cell formation in BMDMs induced by ox-LDL. (A and B) Representative images and Statistical analysis of Oil Red O staining in BMDMs are shown for both A and B (n = 4). (C, D, E, F, G) RBG reduces the ratio of M1 macrophages while increases the ratio of M2 macrophages in BMDM polarization induced by ox-LDL or LPS + IFN-γ (n = 4). (H, I, J, K, L, M) Levels of IL18, IL1β, TNF-α, IL10, TG, and TC in the supernatants of BMDM cultures (n = 8). (N, O, P, Q, R, S, T, U, V) Western blot and statistical analysis was conducted for NLRP3, total ASC protein, ASC oligomerization, NEK7, Caspase-1, Cleaved Caspase-1, Pro IL18, and IL18 (n = 3). (W, a, b, c, d) Representative images and Statistical analysis of Immunofluorescence staining of NLRP3, ASC, and Caspase-1 in BMDMs (n = 3). (X and Y) Representative images of NF-κB immunofluorescence in BMDMs. Ratio of NF-κB fluorescence intensity in the nucleus to the cytoplasm (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns indicates not significant. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Journal: Journal of Advanced Research

    Article Title: Resibufogenin protects against atherosclerosis in ApoE -/- mice through blocking NLRP3 inflammasome assembly

    doi: 10.1016/j.jare.2025.04.029

    Figure Lengend Snippet: RBG enhances foam cell formation in BMDMs induced by ox-LDL. (A and B) Representative images and Statistical analysis of Oil Red O staining in BMDMs are shown for both A and B (n = 4). (C, D, E, F, G) RBG reduces the ratio of M1 macrophages while increases the ratio of M2 macrophages in BMDM polarization induced by ox-LDL or LPS + IFN-γ (n = 4). (H, I, J, K, L, M) Levels of IL18, IL1β, TNF-α, IL10, TG, and TC in the supernatants of BMDM cultures (n = 8). (N, O, P, Q, R, S, T, U, V) Western blot and statistical analysis was conducted for NLRP3, total ASC protein, ASC oligomerization, NEK7, Caspase-1, Cleaved Caspase-1, Pro IL18, and IL18 (n = 3). (W, a, b, c, d) Representative images and Statistical analysis of Immunofluorescence staining of NLRP3, ASC, and Caspase-1 in BMDMs (n = 3). (X and Y) Representative images of NF-κB immunofluorescence in BMDMs. Ratio of NF-κB fluorescence intensity in the nucleus to the cytoplasm (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns indicates not significant. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: Primary antibodies for NLRP3 (ab15101), NF-κB (ab4764), NEK7 (ab10054), ASC (ab67824), Caspase-1 (ab24232), IL18 (ab57058), IL1β (ab31202), CD68 (ab97778), GAPDH (ab2118), and β-ACTIN (ab4970) were obtained from Cell Signaling Technology, Denvers, MA, USA.The antibodies PE Anti-Mouse CD86 (GL-1), APC Anti-Mouse CD206/MMR (C068C2), and PE/Cyanine7 Anti-Mouse F4/80 (CI A3-1) were procured from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

    Techniques: Staining, Western Blot, Immunofluorescence, Fluorescence

    CYS-279 residue of NLRP3 is the binding site for RBG. (A) Sitemap predicted cavity of human NLRP3 protein, along with a covalent site map where spheres represent the CYS covalent reaction site, and the cyan block indicates the pocket cavity. (B) Binding mode of RBG with wild-type human NLRP3 protein. (C) Binding mode of RBG with the R147A mutant of human NLRP3 protein. (D) Variations in the root mean square deviation (RMSD) of the system throughout the molecular dynamics simulation. (E) Temporal variations in energy. (F) RMSD changes over time after 500 ns of extended simulation and for the R147A mutant simulation. (G) SPR response of RBG with purified human NLRP3 protein. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Journal: Journal of Advanced Research

    Article Title: Resibufogenin protects against atherosclerosis in ApoE -/- mice through blocking NLRP3 inflammasome assembly

    doi: 10.1016/j.jare.2025.04.029

    Figure Lengend Snippet: CYS-279 residue of NLRP3 is the binding site for RBG. (A) Sitemap predicted cavity of human NLRP3 protein, along with a covalent site map where spheres represent the CYS covalent reaction site, and the cyan block indicates the pocket cavity. (B) Binding mode of RBG with wild-type human NLRP3 protein. (C) Binding mode of RBG with the R147A mutant of human NLRP3 protein. (D) Variations in the root mean square deviation (RMSD) of the system throughout the molecular dynamics simulation. (E) Temporal variations in energy. (F) RMSD changes over time after 500 ns of extended simulation and for the R147A mutant simulation. (G) SPR response of RBG with purified human NLRP3 protein. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: Primary antibodies for NLRP3 (ab15101), NF-κB (ab4764), NEK7 (ab10054), ASC (ab67824), Caspase-1 (ab24232), IL18 (ab57058), IL1β (ab31202), CD68 (ab97778), GAPDH (ab2118), and β-ACTIN (ab4970) were obtained from Cell Signaling Technology, Denvers, MA, USA.The antibodies PE Anti-Mouse CD86 (GL-1), APC Anti-Mouse CD206/MMR (C068C2), and PE/Cyanine7 Anti-Mouse F4/80 (CI A3-1) were procured from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

    Techniques: Residue, Binding Assay, Blocking Assay, Mutagenesis, Purification

    RBG Inhibits Inflammation and NLRP3 Inflammasome Assembly Induced by LPS + ATP in BMDMs. (A and B) Concentrations of IL18 and IL1β in BMDM culture supernatants (n = 8). (C–H) Relative mRNA expression levels of NLRP3, NEK7, ASC, Caspase-1, IL18, and IL1β were measured in BMDMs (n = 4). (I-Q) Western blot analysis and statistical analysis of NLRP3, ASC total protein, ASC oligomerization, NEK7, Caspase-1, Cleaved Caspase-1, Pro IL18, and IL18 (n = 3). (R, U, V, W, X) Representative immunofluorescence images and statistical analysis of NLRP3, ASC, and Caspase-1 in BMDM (n = 3). (S and T) Representative images of NF-κB immunofluorescence in BMDMs. Ratio of NF-κB relative fluorescence intensity between the nucleus and cytoplasm (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns indicates not significant.

    Journal: Journal of Advanced Research

    Article Title: Resibufogenin protects against atherosclerosis in ApoE -/- mice through blocking NLRP3 inflammasome assembly

    doi: 10.1016/j.jare.2025.04.029

    Figure Lengend Snippet: RBG Inhibits Inflammation and NLRP3 Inflammasome Assembly Induced by LPS + ATP in BMDMs. (A and B) Concentrations of IL18 and IL1β in BMDM culture supernatants (n = 8). (C–H) Relative mRNA expression levels of NLRP3, NEK7, ASC, Caspase-1, IL18, and IL1β were measured in BMDMs (n = 4). (I-Q) Western blot analysis and statistical analysis of NLRP3, ASC total protein, ASC oligomerization, NEK7, Caspase-1, Cleaved Caspase-1, Pro IL18, and IL18 (n = 3). (R, U, V, W, X) Representative immunofluorescence images and statistical analysis of NLRP3, ASC, and Caspase-1 in BMDM (n = 3). (S and T) Representative images of NF-κB immunofluorescence in BMDMs. Ratio of NF-κB relative fluorescence intensity between the nucleus and cytoplasm (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns indicates not significant.

    Article Snippet: Primary antibodies for NLRP3 (ab15101), NF-κB (ab4764), NEK7 (ab10054), ASC (ab67824), Caspase-1 (ab24232), IL18 (ab57058), IL1β (ab31202), CD68 (ab97778), GAPDH (ab2118), and β-ACTIN (ab4970) were obtained from Cell Signaling Technology, Denvers, MA, USA.The antibodies PE Anti-Mouse CD86 (GL-1), APC Anti-Mouse CD206/MMR (C068C2), and PE/Cyanine7 Anti-Mouse F4/80 (CI A3-1) were procured from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

    Techniques: Expressing, Western Blot, Immunofluorescence, Fluorescence

    The Inhibitory Effect of RBG on LPS + ATP Induced Inflammatory Responses in BMDMs Depends on NLRP3. (A, B, C) Concentrations of IL18, IL1β, and Caspase-1 in the culture supernatants of NLRP3-overexpressing BMDMs (n = 8). (D, E, F) Analysis of total protein and ASC oligomerization in NLRP3-overexpressing BMDMs (n = 3). (G, H, J, K) Representative immunofluorescence images and statistical analysis of NLRP3, ASC, and Caspase-1 in NLRP3-overexpressing BMDMs (n = 3). (L and M) Representative images of NF-κB immunofluorescence in NLRP3-overexpressing BMDMs. Ratio of fluorescence intensity of NF-κB in the nucleus to that in the cytoplasm (n = 3). Concentrations of IL18, IL1β, and Caspase-1 in the supernatants of NLRP3-knockdown BMDMs (n = 8). (Q, R, S) Total protein and oligomerization levels of ASC in NLRP3-knockdown BMDMs (n = 3). Representative immunofluorescence images and statistical analysis of NLRP3, ASC, and Caspase-1 staining in NLRP3-knockdown BMDMs (n = 3). (Y and Z) Representative images of NF-κB immunofluorescence in NLRP3-knockdown BMDMs. Ratio of NF-κB relative fluorescence intensity between the nucleus and cytoplasm (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns indicates not significant.

    Journal: Journal of Advanced Research

    Article Title: Resibufogenin protects against atherosclerosis in ApoE -/- mice through blocking NLRP3 inflammasome assembly

    doi: 10.1016/j.jare.2025.04.029

    Figure Lengend Snippet: The Inhibitory Effect of RBG on LPS + ATP Induced Inflammatory Responses in BMDMs Depends on NLRP3. (A, B, C) Concentrations of IL18, IL1β, and Caspase-1 in the culture supernatants of NLRP3-overexpressing BMDMs (n = 8). (D, E, F) Analysis of total protein and ASC oligomerization in NLRP3-overexpressing BMDMs (n = 3). (G, H, J, K) Representative immunofluorescence images and statistical analysis of NLRP3, ASC, and Caspase-1 in NLRP3-overexpressing BMDMs (n = 3). (L and M) Representative images of NF-κB immunofluorescence in NLRP3-overexpressing BMDMs. Ratio of fluorescence intensity of NF-κB in the nucleus to that in the cytoplasm (n = 3). Concentrations of IL18, IL1β, and Caspase-1 in the supernatants of NLRP3-knockdown BMDMs (n = 8). (Q, R, S) Total protein and oligomerization levels of ASC in NLRP3-knockdown BMDMs (n = 3). Representative immunofluorescence images and statistical analysis of NLRP3, ASC, and Caspase-1 staining in NLRP3-knockdown BMDMs (n = 3). (Y and Z) Representative images of NF-κB immunofluorescence in NLRP3-knockdown BMDMs. Ratio of NF-κB relative fluorescence intensity between the nucleus and cytoplasm (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns indicates not significant.

    Article Snippet: Primary antibodies for NLRP3 (ab15101), NF-κB (ab4764), NEK7 (ab10054), ASC (ab67824), Caspase-1 (ab24232), IL18 (ab57058), IL1β (ab31202), CD68 (ab97778), GAPDH (ab2118), and β-ACTIN (ab4970) were obtained from Cell Signaling Technology, Denvers, MA, USA.The antibodies PE Anti-Mouse CD86 (GL-1), APC Anti-Mouse CD206/MMR (C068C2), and PE/Cyanine7 Anti-Mouse F4/80 (CI A3-1) were procured from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

    Techniques: Immunofluorescence, Fluorescence, Knockdown, Staining

    The Inhibitory Effect of RBG on Ox-LDL Induced Inflammatory Responses and Foam Cell Formation in BMDMs Depends on NLRP3. (A) Representative images of Oil Red O staining in NLRP3-overexpressing BMDMs (n = 6). (B, C, D, E, F, G) Concentrations of IL18, IL1β, TNF-α, IL10, TG, and TC in the culture supernatants of NLRP3-overexpressing BMDMs (n = 8). (H, I, J, K, L) Representative immunofluorescence images and statistical analysis of NLRP3, ASC, and Caspase-1 in NLRP3-overexpressing BMDMs (n = 3). (M and N) Representative images of NF-κB immunofluorescence in NLRP3-overexpressing BMDMs. Ratio of relative fluorescence intensity of NF-κB in the nucleus to that in the cytoplasm (n = 3). (O) Representative images of Oil Red O staining in NLRP3-knockdown BMDMs (n = 6). (P, Q, R, S, T, U) Concentrations of IL18, IL1β, TNF-α, IL10, TG, and TC in the culture supernatants of NLRP3-knockdown BMDMs (n = 8). (V, W, X, Y Z) Representative immunofluorescence images and statistical analysis of NLRP3, ASC, and Caspase1 in NLRP3-knockdown BMDMs (n = 3). (a and b) Representative images of NF-κB immunofluorescence in NLRP3-knockdown BMDMs. Ratio of relative fluorescence intensity of NF-κB in the nucleus to that in the cytoplasm (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 and ns: Not significant. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Journal: Journal of Advanced Research

    Article Title: Resibufogenin protects against atherosclerosis in ApoE -/- mice through blocking NLRP3 inflammasome assembly

    doi: 10.1016/j.jare.2025.04.029

    Figure Lengend Snippet: The Inhibitory Effect of RBG on Ox-LDL Induced Inflammatory Responses and Foam Cell Formation in BMDMs Depends on NLRP3. (A) Representative images of Oil Red O staining in NLRP3-overexpressing BMDMs (n = 6). (B, C, D, E, F, G) Concentrations of IL18, IL1β, TNF-α, IL10, TG, and TC in the culture supernatants of NLRP3-overexpressing BMDMs (n = 8). (H, I, J, K, L) Representative immunofluorescence images and statistical analysis of NLRP3, ASC, and Caspase-1 in NLRP3-overexpressing BMDMs (n = 3). (M and N) Representative images of NF-κB immunofluorescence in NLRP3-overexpressing BMDMs. Ratio of relative fluorescence intensity of NF-κB in the nucleus to that in the cytoplasm (n = 3). (O) Representative images of Oil Red O staining in NLRP3-knockdown BMDMs (n = 6). (P, Q, R, S, T, U) Concentrations of IL18, IL1β, TNF-α, IL10, TG, and TC in the culture supernatants of NLRP3-knockdown BMDMs (n = 8). (V, W, X, Y Z) Representative immunofluorescence images and statistical analysis of NLRP3, ASC, and Caspase1 in NLRP3-knockdown BMDMs (n = 3). (a and b) Representative images of NF-κB immunofluorescence in NLRP3-knockdown BMDMs. Ratio of relative fluorescence intensity of NF-κB in the nucleus to that in the cytoplasm (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 and ns: Not significant. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: Primary antibodies for NLRP3 (ab15101), NF-κB (ab4764), NEK7 (ab10054), ASC (ab67824), Caspase-1 (ab24232), IL18 (ab57058), IL1β (ab31202), CD68 (ab97778), GAPDH (ab2118), and β-ACTIN (ab4970) were obtained from Cell Signaling Technology, Denvers, MA, USA.The antibodies PE Anti-Mouse CD86 (GL-1), APC Anti-Mouse CD206/MMR (C068C2), and PE/Cyanine7 Anti-Mouse F4/80 (CI A3-1) were procured from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

    Techniques: Staining, Immunofluorescence, Fluorescence, Knockdown

    The beneficial impact of RBG on AS caused by HFD in ApoE -/- mice relies on NLRP3. (A) Construction of vascular-specific NLRP3 knockdown in ApoE -/- mice, model replication, and treatment protocol. (B) Body weight-time curve of mice (n = 10). (C, D) Representative images and quantification of lesion area of Oil Red O staining of the aorta (n = 3). (E-L) Serum levels of TG, TC, LDL-C, HDL-C, IL18, IL1β, TNF-α, and IL10 were measured (n = 10). (M−P) Representative images and quantification of lesion area of HE, MASSON, and Oil Red O staining (n = 3). (Q) Comparison of mouse body weight at the end of the experiment. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns indicates not significant. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Journal: Journal of Advanced Research

    Article Title: Resibufogenin protects against atherosclerosis in ApoE -/- mice through blocking NLRP3 inflammasome assembly

    doi: 10.1016/j.jare.2025.04.029

    Figure Lengend Snippet: The beneficial impact of RBG on AS caused by HFD in ApoE -/- mice relies on NLRP3. (A) Construction of vascular-specific NLRP3 knockdown in ApoE -/- mice, model replication, and treatment protocol. (B) Body weight-time curve of mice (n = 10). (C, D) Representative images and quantification of lesion area of Oil Red O staining of the aorta (n = 3). (E-L) Serum levels of TG, TC, LDL-C, HDL-C, IL18, IL1β, TNF-α, and IL10 were measured (n = 10). (M−P) Representative images and quantification of lesion area of HE, MASSON, and Oil Red O staining (n = 3). (Q) Comparison of mouse body weight at the end of the experiment. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns indicates not significant. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: Primary antibodies for NLRP3 (ab15101), NF-κB (ab4764), NEK7 (ab10054), ASC (ab67824), Caspase-1 (ab24232), IL18 (ab57058), IL1β (ab31202), CD68 (ab97778), GAPDH (ab2118), and β-ACTIN (ab4970) were obtained from Cell Signaling Technology, Denvers, MA, USA.The antibodies PE Anti-Mouse CD86 (GL-1), APC Anti-Mouse CD206/MMR (C068C2), and PE/Cyanine7 Anti-Mouse F4/80 (CI A3-1) were procured from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

    Techniques: Knockdown, Staining, Comparison

    Differences in pyroptosis induced by COM crystals of different sizes (A) Caspase-1/PI double staining flow cytometry quantitative analysis. (B) Caspase-1/PI/Hoechst 33342 triple staining confocal observation. Scale bars, 50 μm. (C) NLRP3 immunofluorescence map. Scale bars, 100 μm. (D) Quantitative histogram of pyroptosis; n = 3; mean ± SEM. (E) Quantitative bar graph of NLRP3; n = 3; mean ± SEM. NC: normal control. COM crystals were incubated with HK-2 cells for 48 h. Compared with NC group, ∗ p < 0.05; ∗∗ p < 0.01. (A) flow graph is divided into four regions (Q1, Q2, Q3, and Q4), of which Q1 is PI high signal area and Caspase-1 low signal area, representing apoptotic necrotic cells. Q2 is the PI high signal region and Caspase-1 high signal region, which represents the late pyroptosis cells. Q3 is the PI low signal area and Caspase-1 high signal area, which represents the early pyroptosis cells (Caspase-1 is actively expressed). Q4 is the PI hypointense region and Caspase-1 hypointense region, representing normal cells. Q1+ Q2 refers to dead cells, and Q2+ Q3 refers to pyroptosis cells. (B) shows the presence of four types of cells: the first type is the normal cell (indicated by the orange arrow), which corresponds to the Q4 region cells in (A); the second type was apoptotic or necrotic cells (indicated by white arrow), namely Q1 region cells. The third type was the late pyroptosis cells (yellow arrow), which were Q2 cells. The fourth type is the cells in the early stage of pyroptosis (purple arrow), which is the Q3 region cells. Orange arrows refer to normal cells, white arrows to apoptotic necrotic cells, yellow arrows to late pyroptosis cells, and purple arrows to early pyroptosis cells.

    Journal: iScience

    Article Title: Size-dependent pyroptosis induction by calcium oxalate monohydrate crystals in HK-2 cells

    doi: 10.1016/j.isci.2025.114459

    Figure Lengend Snippet: Differences in pyroptosis induced by COM crystals of different sizes (A) Caspase-1/PI double staining flow cytometry quantitative analysis. (B) Caspase-1/PI/Hoechst 33342 triple staining confocal observation. Scale bars, 50 μm. (C) NLRP3 immunofluorescence map. Scale bars, 100 μm. (D) Quantitative histogram of pyroptosis; n = 3; mean ± SEM. (E) Quantitative bar graph of NLRP3; n = 3; mean ± SEM. NC: normal control. COM crystals were incubated with HK-2 cells for 48 h. Compared with NC group, ∗ p < 0.05; ∗∗ p < 0.01. (A) flow graph is divided into four regions (Q1, Q2, Q3, and Q4), of which Q1 is PI high signal area and Caspase-1 low signal area, representing apoptotic necrotic cells. Q2 is the PI high signal region and Caspase-1 high signal region, which represents the late pyroptosis cells. Q3 is the PI low signal area and Caspase-1 high signal area, which represents the early pyroptosis cells (Caspase-1 is actively expressed). Q4 is the PI hypointense region and Caspase-1 hypointense region, representing normal cells. Q1+ Q2 refers to dead cells, and Q2+ Q3 refers to pyroptosis cells. (B) shows the presence of four types of cells: the first type is the normal cell (indicated by the orange arrow), which corresponds to the Q4 region cells in (A); the second type was apoptotic or necrotic cells (indicated by white arrow), namely Q1 region cells. The third type was the late pyroptosis cells (yellow arrow), which were Q2 cells. The fourth type is the cells in the early stage of pyroptosis (purple arrow), which is the Q3 region cells. Orange arrows refer to normal cells, white arrows to apoptotic necrotic cells, yellow arrows to late pyroptosis cells, and purple arrows to early pyroptosis cells.

    Article Snippet: After reaching the incubation time, the culture medium was removed, washed twice with PBS, fixed with paraformaldehyde for 15 min, blocked with sheep serum for 20 min, and then incubated with NLRP3 primary antibody (1:100) (Proteintech, Cat No. 30109-1-AP) at 4°C overnight.

    Techniques: Double Staining, Flow Cytometry, Staining, Immunofluorescence, Control, Incubation

    Level of proteins in pyroptosis signaling pathway induced by COM crystals of different sizes (A) protein bands of NLRP3, cleaved caspase-1 p20, IL-18, and IL-1β. (B–E) Quantitative analysis of NLRP3, cleaved caspase-1 p20, IL-18, and IL-1β, respectively. n = 3; mean ± SEM. NC: normal control. COM crystals were incubated with HK-2 cells for 48 h.∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

    Journal: iScience

    Article Title: Size-dependent pyroptosis induction by calcium oxalate monohydrate crystals in HK-2 cells

    doi: 10.1016/j.isci.2025.114459

    Figure Lengend Snippet: Level of proteins in pyroptosis signaling pathway induced by COM crystals of different sizes (A) protein bands of NLRP3, cleaved caspase-1 p20, IL-18, and IL-1β. (B–E) Quantitative analysis of NLRP3, cleaved caspase-1 p20, IL-18, and IL-1β, respectively. n = 3; mean ± SEM. NC: normal control. COM crystals were incubated with HK-2 cells for 48 h.∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

    Article Snippet: After reaching the incubation time, the culture medium was removed, washed twice with PBS, fixed with paraformaldehyde for 15 min, blocked with sheep serum for 20 min, and then incubated with NLRP3 primary antibody (1:100) (Proteintech, Cat No. 30109-1-AP) at 4°C overnight.

    Techniques: Control, Incubation