cleaved gasdermin d  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc cleaved gasdermin d
    Cleaved Gasdermin D, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    10137s  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc 10137s
    Reagent and resources.
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    1) Product Images from "An IBD-associated pathobiont synergises with NSAID to promote colitis which is blocked by NLRP3 inflammasome and Caspase-8 inhibitors"

    Article Title: An IBD-associated pathobiont synergises with NSAID to promote colitis which is blocked by NLRP3 inflammasome and Caspase-8 inhibitors

    Journal: Gut Microbes

    doi: 10.1080/19490976.2022.2163838

    Reagent and resources.
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    Techniques Used: Blocking Assay, Recombinant, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Biomarker Assay, Sequencing, Software

    cleaved gasdermin d  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc cleaved gasdermin d
    Cleaved Gasdermin D, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit anti mouse cleaved caspase-9  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti mouse cleaved caspase-9
    Rabbit Anti Mouse Cleaved Caspase 9, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit anti gasdermin d l60  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti gasdermin d l60
    Reagents List.
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    1) Product Images from "Complement membrane attack complex is an immunometabolic regulator of NLRP3 activation and IL-18 secretion in human macrophages"

    Article Title: Complement membrane attack complex is an immunometabolic regulator of NLRP3 activation and IL-18 secretion in human macrophages

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2022.918551

    Reagents List.
    Figure Legend Snippet: Reagents List.

    Techniques Used: Purification, Recombinant, In Vivo, Generated, Enzyme-linked Immunosorbent Assay, Sequencing, Software

    cleaved gsdmd  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc cleaved gsdmd
    Cleaved Gsdmd, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    monoclonal rabbit anti cleaved gsdmd asp276  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc monoclonal rabbit anti cleaved gsdmd asp276
    Monoclonal Rabbit Anti Cleaved Gsdmd Asp276, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit anti cleaved gsdmd  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti cleaved gsdmd
    Pyroptosis in the murine TBI model. To assess the expression of pyroptosis-related proteins, TBI mice were sacrificed and brain tissues were removed at 12 h, 24 h, 48 h and 72 h post-TBI surgery. a Representative protein bands and corresponding grayscale values of <t>NLRP3,</t> <t>pro-Caspase-1,</t> Caspase-1 p20, <t>GSDMD,</t> and cleaved-GSDMD performed by western blotting. b – d Representative double immunofluorescence staining photographs and quantification of Iba-1 (green) with NLRP3 (red), Caspase-1 p20 (red), or GSDMD (red) in the injured cortex. Scale bar = 50 μm. All data are represented as means ± SD (n = 5 mice/group) and compared by t -test. *p < 0.05, ** p < 0.01, *** p < 0.001 versus sham group
    Rabbit Anti Cleaved Gsdmd, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Mesenchymal stem cells protect against TBI-induced pyroptosis in vivo and in vitro through TSG-6"

    Article Title: Mesenchymal stem cells protect against TBI-induced pyroptosis in vivo and in vitro through TSG-6

    Journal: Cell Communication and Signaling : CCS

    doi: 10.1186/s12964-022-00931-2

    Pyroptosis in the murine TBI model. To assess the expression of pyroptosis-related proteins, TBI mice were sacrificed and brain tissues were removed at 12 h, 24 h, 48 h and 72 h post-TBI surgery. a Representative protein bands and corresponding grayscale values of NLRP3, pro-Caspase-1, Caspase-1 p20, GSDMD, and cleaved-GSDMD performed by western blotting. b – d Representative double immunofluorescence staining photographs and quantification of Iba-1 (green) with NLRP3 (red), Caspase-1 p20 (red), or GSDMD (red) in the injured cortex. Scale bar = 50 μm. All data are represented as means ± SD (n = 5 mice/group) and compared by t -test. *p < 0.05, ** p < 0.01, *** p < 0.001 versus sham group
    Figure Legend Snippet: Pyroptosis in the murine TBI model. To assess the expression of pyroptosis-related proteins, TBI mice were sacrificed and brain tissues were removed at 12 h, 24 h, 48 h and 72 h post-TBI surgery. a Representative protein bands and corresponding grayscale values of NLRP3, pro-Caspase-1, Caspase-1 p20, GSDMD, and cleaved-GSDMD performed by western blotting. b – d Representative double immunofluorescence staining photographs and quantification of Iba-1 (green) with NLRP3 (red), Caspase-1 p20 (red), or GSDMD (red) in the injured cortex. Scale bar = 50 μm. All data are represented as means ± SD (n = 5 mice/group) and compared by t -test. *p < 0.05, ** p < 0.01, *** p < 0.001 versus sham group

    Techniques Used: Expressing, Western Blot, Double Immunofluorescence Staining

    hUMSCs downregulated the NLRP3/Caspase-1/GSDMD signaling pathway and reduced microglia pyroptosis in TBI mice via TSG-6. TBI mice were intracerebroventricularly injected with 10 μL of PBS, 0.5 µg rmTSG-6 or 1 µg rmTSG-6, or 2.5 × 10 5 hUMSCs, sh-TSG-6 hUMSCs, sh-NC hUMSCs, in 10 μL of PBS at 4 h after TBI surgery. Then TBI mice were sacrificed and brain tissues were removed at 48 h post-TBI surgery. a Representative protein bands and corresponding grayscale values of NLRP3, pro-Caspase-1, Caspase-1 p20, GSDMD, and cleaved-GSDMD performed by western blotting. b – d Representative double immunofluorescence staining photographs and quantification of Iba-1 (green) with NLRP3 (red), Caspase-1 p20 (red), or GSDMD (red) in the injured cortex. Scale bar = 50 μm. All data are represented as means ± SD (n = 6 mice/group) and compared by one-way ANOVA, followed by the Tukey’s post hoc test. * p < 0.05, ** p < 0.01, versus TBI group; # p < 0.05, sh-TSG-6 hUMSC group versus sh-NC hUMSC group
    Figure Legend Snippet: hUMSCs downregulated the NLRP3/Caspase-1/GSDMD signaling pathway and reduced microglia pyroptosis in TBI mice via TSG-6. TBI mice were intracerebroventricularly injected with 10 μL of PBS, 0.5 µg rmTSG-6 or 1 µg rmTSG-6, or 2.5 × 10 5 hUMSCs, sh-TSG-6 hUMSCs, sh-NC hUMSCs, in 10 μL of PBS at 4 h after TBI surgery. Then TBI mice were sacrificed and brain tissues were removed at 48 h post-TBI surgery. a Representative protein bands and corresponding grayscale values of NLRP3, pro-Caspase-1, Caspase-1 p20, GSDMD, and cleaved-GSDMD performed by western blotting. b – d Representative double immunofluorescence staining photographs and quantification of Iba-1 (green) with NLRP3 (red), Caspase-1 p20 (red), or GSDMD (red) in the injured cortex. Scale bar = 50 μm. All data are represented as means ± SD (n = 6 mice/group) and compared by one-way ANOVA, followed by the Tukey’s post hoc test. * p < 0.05, ** p < 0.01, versus TBI group; # p < 0.05, sh-TSG-6 hUMSC group versus sh-NC hUMSC group

    Techniques Used: Injection, Western Blot, Double Immunofluorescence Staining

    hUMSCs reduced LPS + ATP-induced BV2 microglia pyroptosis by secreting TSG-6. BV2 cells were co-cultured with hUMSCs, sh-TSG-6 hUMSCs, or sh-NC hUMSCs activated by TNF-α and different concentration of rmTSG-6 (50 ng/mL, 100 ng/mL, 200 ng/mL) and treated with or without LPS (1 µg/mL) + ATP (5 mM) for 12 h. a Representative photomicrographs and quantification of double staining with PI (red) and DAPI (blue) were captured by confocal microscopy. b Caspase-1 activity and the release of LDH in the supernatant was assessed using Caspase-1 activity and LDH activity detection kits. c The protein levels of IL-1β, pro IL-18, and TNF-α were evaluated by ELISA. d Representative protein bands and corresponding grayscale values of NLRP3, pro-Caspase-1, Caspase-1 p20, GSDMD and cleaved-GSDMD were performed by western blotting. Scale bar = 100 μm. All data are represented as means ± SD of at least 3 independent experiments and compared by one-way ANOVA, followed by the Tukey’s post hoc test. * p < 0.05, ** p < 0.01, versus LPS + ATP group; # p < 0.05, sh-TSG-6 hUMSC group versus sh-NC hUMSC group
    Figure Legend Snippet: hUMSCs reduced LPS + ATP-induced BV2 microglia pyroptosis by secreting TSG-6. BV2 cells were co-cultured with hUMSCs, sh-TSG-6 hUMSCs, or sh-NC hUMSCs activated by TNF-α and different concentration of rmTSG-6 (50 ng/mL, 100 ng/mL, 200 ng/mL) and treated with or without LPS (1 µg/mL) + ATP (5 mM) for 12 h. a Representative photomicrographs and quantification of double staining with PI (red) and DAPI (blue) were captured by confocal microscopy. b Caspase-1 activity and the release of LDH in the supernatant was assessed using Caspase-1 activity and LDH activity detection kits. c The protein levels of IL-1β, pro IL-18, and TNF-α were evaluated by ELISA. d Representative protein bands and corresponding grayscale values of NLRP3, pro-Caspase-1, Caspase-1 p20, GSDMD and cleaved-GSDMD were performed by western blotting. Scale bar = 100 μm. All data are represented as means ± SD of at least 3 independent experiments and compared by one-way ANOVA, followed by the Tukey’s post hoc test. * p < 0.05, ** p < 0.01, versus LPS + ATP group; # p < 0.05, sh-TSG-6 hUMSC group versus sh-NC hUMSC group

    Techniques Used: Cell Culture, Concentration Assay, Double Staining, Confocal Microscopy, Activity Assay, Enzyme-linked Immunosorbent Assay, Western Blot

    gasdermin d n gsdmd n  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc gasdermin d n gsdmd n
    Gasdermin D N Gsdmd N, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti cleaved gasdermin  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti cleaved gasdermin
    <t>Gasdermin</t> d -deficient mice ( Gsdmd -/- ) have reduced pulmonary inflammation and remodeling after acute smoke cigarette (CS) exposure. WT and Gsdmd -/- mice were exposed to CS or Air during 4 days. Total cells (A) , macrophages (B) and neutrophils (C) counts and MPO level in BAL (D) are shown. CXCL1 (E) , CXCL5 (F) , CXCL15 (G) and IL-1β (H) levels were measured in BAL. MMP-9 (I) and TIMP-1 (J) remodeling factor levels were measured in BAL. In lungs, levels of IL-1β (K) , MMP-9 (L) and TIMP-1 (M) were also measured. All parameters decreased in Gsdmd -/- mice exposed to CS as compared to CS WT mice. Data are representative of four experiments and are expressed as mean values ± SEM (n= 4-6 mice per group, *p < 0.05, **p < 0.01, ns, non significant, using a Mann Whitney test).
    Anti Cleaved Gasdermin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Cigarette smoke-induced gasdermin D activation in bronchoalveolar macrophages and bronchial epithelial cells dependently on NLRP3"

    Article Title: Cigarette smoke-induced gasdermin D activation in bronchoalveolar macrophages and bronchial epithelial cells dependently on NLRP3

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2022.918507

    Gasdermin d -deficient mice ( Gsdmd -/- ) have reduced pulmonary inflammation and remodeling after acute smoke cigarette (CS) exposure. WT and Gsdmd -/- mice were exposed to CS or Air during 4 days. Total cells (A) , macrophages (B) and neutrophils (C) counts and MPO level in BAL (D) are shown. CXCL1 (E) , CXCL5 (F) , CXCL15 (G) and IL-1β (H) levels were measured in BAL. MMP-9 (I) and TIMP-1 (J) remodeling factor levels were measured in BAL. In lungs, levels of IL-1β (K) , MMP-9 (L) and TIMP-1 (M) were also measured. All parameters decreased in Gsdmd -/- mice exposed to CS as compared to CS WT mice. Data are representative of four experiments and are expressed as mean values ± SEM (n= 4-6 mice per group, *p < 0.05, **p < 0.01, ns, non significant, using a Mann Whitney test).
    Figure Legend Snippet: Gasdermin d -deficient mice ( Gsdmd -/- ) have reduced pulmonary inflammation and remodeling after acute smoke cigarette (CS) exposure. WT and Gsdmd -/- mice were exposed to CS or Air during 4 days. Total cells (A) , macrophages (B) and neutrophils (C) counts and MPO level in BAL (D) are shown. CXCL1 (E) , CXCL5 (F) , CXCL15 (G) and IL-1β (H) levels were measured in BAL. MMP-9 (I) and TIMP-1 (J) remodeling factor levels were measured in BAL. In lungs, levels of IL-1β (K) , MMP-9 (L) and TIMP-1 (M) were also measured. All parameters decreased in Gsdmd -/- mice exposed to CS as compared to CS WT mice. Data are representative of four experiments and are expressed as mean values ± SEM (n= 4-6 mice per group, *p < 0.05, **p < 0.01, ns, non significant, using a Mann Whitney test).

    Techniques Used: MANN-WHITNEY

    mouse anti cleaved gasdermin d  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc mouse anti cleaved gasdermin d
    <t>Gasdermin</t> <t>d</t> -deficient mice ( Gsdmd -/- ) have reduced pulmonary inflammation and remodeling after acute smoke cigarette (CS) exposure. WT and Gsdmd -/- mice were exposed to CS or Air during 4 days. Total cells (A) , macrophages (B) and neutrophils (C) counts and MPO level in BAL (D) are shown. CXCL1 (E) , CXCL5 (F) , CXCL15 (G) and IL-1β (H) levels were measured in BAL. MMP-9 (I) and TIMP-1 (J) remodeling factor levels were measured in BAL. In lungs, levels of IL-1β (K) , MMP-9 (L) and TIMP-1 (M) were also measured. All parameters decreased in Gsdmd -/- mice exposed to CS as compared to CS WT mice. Data are representative of four experiments and are expressed as mean values ± SEM (n= 4-6 mice per group, *p < 0.05, **p < 0.01, ns, non significant, using a Mann Whitney test).
    Mouse Anti Cleaved Gasdermin D, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti cleaved gasdermin d/product/Cell Signaling Technology Inc
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    1) Product Images from "Cigarette smoke-induced gasdermin D activation in bronchoalveolar macrophages and bronchial epithelial cells dependently on NLRP3"

    Article Title: Cigarette smoke-induced gasdermin D activation in bronchoalveolar macrophages and bronchial epithelial cells dependently on NLRP3

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2022.918507

    Gasdermin d -deficient mice ( Gsdmd -/- ) have reduced pulmonary inflammation and remodeling after acute smoke cigarette (CS) exposure. WT and Gsdmd -/- mice were exposed to CS or Air during 4 days. Total cells (A) , macrophages (B) and neutrophils (C) counts and MPO level in BAL (D) are shown. CXCL1 (E) , CXCL5 (F) , CXCL15 (G) and IL-1β (H) levels were measured in BAL. MMP-9 (I) and TIMP-1 (J) remodeling factor levels were measured in BAL. In lungs, levels of IL-1β (K) , MMP-9 (L) and TIMP-1 (M) were also measured. All parameters decreased in Gsdmd -/- mice exposed to CS as compared to CS WT mice. Data are representative of four experiments and are expressed as mean values ± SEM (n= 4-6 mice per group, *p < 0.05, **p < 0.01, ns, non significant, using a Mann Whitney test).
    Figure Legend Snippet: Gasdermin d -deficient mice ( Gsdmd -/- ) have reduced pulmonary inflammation and remodeling after acute smoke cigarette (CS) exposure. WT and Gsdmd -/- mice were exposed to CS or Air during 4 days. Total cells (A) , macrophages (B) and neutrophils (C) counts and MPO level in BAL (D) are shown. CXCL1 (E) , CXCL5 (F) , CXCL15 (G) and IL-1β (H) levels were measured in BAL. MMP-9 (I) and TIMP-1 (J) remodeling factor levels were measured in BAL. In lungs, levels of IL-1β (K) , MMP-9 (L) and TIMP-1 (M) were also measured. All parameters decreased in Gsdmd -/- mice exposed to CS as compared to CS WT mice. Data are representative of four experiments and are expressed as mean values ± SEM (n= 4-6 mice per group, *p < 0.05, **p < 0.01, ns, non significant, using a Mann Whitney test).

    Techniques Used: MANN-WHITNEY

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    Pyroptosis in the murine TBI model. To assess the expression of pyroptosis-related proteins, TBI mice were sacrificed and brain tissues were removed at 12 h, 24 h, 48 h and 72 h post-TBI surgery. a Representative protein bands and corresponding grayscale values of <t>NLRP3,</t> <t>pro-Caspase-1,</t> Caspase-1 p20, <t>GSDMD,</t> and cleaved-GSDMD performed by western blotting. b – d Representative double immunofluorescence staining photographs and quantification of Iba-1 (green) with NLRP3 (red), Caspase-1 p20 (red), or GSDMD (red) in the injured cortex. Scale bar = 50 μm. All data are represented as means ± SD (n = 5 mice/group) and compared by t -test. *p < 0.05, ** p < 0.01, *** p < 0.001 versus sham group
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    Pyroptosis in the murine TBI model. To assess the expression of pyroptosis-related proteins, TBI mice were sacrificed and brain tissues were removed at 12 h, 24 h, 48 h and 72 h post-TBI surgery. a Representative protein bands and corresponding grayscale values of <t>NLRP3,</t> <t>pro-Caspase-1,</t> Caspase-1 p20, <t>GSDMD,</t> and cleaved-GSDMD performed by western blotting. b – d Representative double immunofluorescence staining photographs and quantification of Iba-1 (green) with NLRP3 (red), Caspase-1 p20 (red), or GSDMD (red) in the injured cortex. Scale bar = 50 μm. All data are represented as means ± SD (n = 5 mice/group) and compared by t -test. *p < 0.05, ** p < 0.01, *** p < 0.001 versus sham group
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    Image Search Results


    Reagent and resources.

    Journal: Gut Microbes

    Article Title: An IBD-associated pathobiont synergises with NSAID to promote colitis which is blocked by NLRP3 inflammasome and Caspase-8 inhibitors

    doi: 10.1080/19490976.2022.2163838

    Figure Lengend Snippet: Reagent and resources.

    Article Snippet: Rabbit monoclonal cleaved gasdermin D (GSDMD) , Cell signaling technology , Cat# 10137S.

    Techniques: Blocking Assay, Recombinant, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Biomarker Assay, Sequencing, Software

    Reagents List.

    Journal: Frontiers in Immunology

    Article Title: Complement membrane attack complex is an immunometabolic regulator of NLRP3 activation and IL-18 secretion in human macrophages

    doi: 10.3389/fimmu.2022.918551

    Figure Lengend Snippet: Reagents List.

    Article Snippet: Rabbit anti-Gasdermin D L60 , Cell Signalling , 93709.

    Techniques: Purification, Recombinant, In Vivo, Generated, Enzyme-linked Immunosorbent Assay, Sequencing, Software

    Pyroptosis in the murine TBI model. To assess the expression of pyroptosis-related proteins, TBI mice were sacrificed and brain tissues were removed at 12 h, 24 h, 48 h and 72 h post-TBI surgery. a Representative protein bands and corresponding grayscale values of NLRP3, pro-Caspase-1, Caspase-1 p20, GSDMD, and cleaved-GSDMD performed by western blotting. b – d Representative double immunofluorescence staining photographs and quantification of Iba-1 (green) with NLRP3 (red), Caspase-1 p20 (red), or GSDMD (red) in the injured cortex. Scale bar = 50 μm. All data are represented as means ± SD (n = 5 mice/group) and compared by t -test. *p < 0.05, ** p < 0.01, *** p < 0.001 versus sham group

    Journal: Cell Communication and Signaling : CCS

    Article Title: Mesenchymal stem cells protect against TBI-induced pyroptosis in vivo and in vitro through TSG-6

    doi: 10.1186/s12964-022-00931-2

    Figure Lengend Snippet: Pyroptosis in the murine TBI model. To assess the expression of pyroptosis-related proteins, TBI mice were sacrificed and brain tissues were removed at 12 h, 24 h, 48 h and 72 h post-TBI surgery. a Representative protein bands and corresponding grayscale values of NLRP3, pro-Caspase-1, Caspase-1 p20, GSDMD, and cleaved-GSDMD performed by western blotting. b – d Representative double immunofluorescence staining photographs and quantification of Iba-1 (green) with NLRP3 (red), Caspase-1 p20 (red), or GSDMD (red) in the injured cortex. Scale bar = 50 μm. All data are represented as means ± SD (n = 5 mice/group) and compared by t -test. *p < 0.05, ** p < 0.01, *** p < 0.001 versus sham group

    Article Snippet: The membranes were blocked for 2 h at 37 °C with 5% non-fat milk and 0.05% Tween-20, then incubated overnight at 4 °C with Mouse anti-GAPDH (60004-1-Ig, Proteintech, 1:5000), rabbit anti-NLRP3 (#15101, CST, 1:1000), rabbit anti-pro-Caspase-1 (AF5418, Affinity, 1:1000), rabbit anti-Caspase-1 p20 (AF4005, Affinity, 1:1000), rabbit anti-GSDMD (ab219800, Abcam, 1:1000), and rabbit anti-cleaved-GSDMD (#10137, CST, 1:1000).

    Techniques: Expressing, Western Blot, Double Immunofluorescence Staining

    hUMSCs downregulated the NLRP3/Caspase-1/GSDMD signaling pathway and reduced microglia pyroptosis in TBI mice via TSG-6. TBI mice were intracerebroventricularly injected with 10 μL of PBS, 0.5 µg rmTSG-6 or 1 µg rmTSG-6, or 2.5 × 10 5 hUMSCs, sh-TSG-6 hUMSCs, sh-NC hUMSCs, in 10 μL of PBS at 4 h after TBI surgery. Then TBI mice were sacrificed and brain tissues were removed at 48 h post-TBI surgery. a Representative protein bands and corresponding grayscale values of NLRP3, pro-Caspase-1, Caspase-1 p20, GSDMD, and cleaved-GSDMD performed by western blotting. b – d Representative double immunofluorescence staining photographs and quantification of Iba-1 (green) with NLRP3 (red), Caspase-1 p20 (red), or GSDMD (red) in the injured cortex. Scale bar = 50 μm. All data are represented as means ± SD (n = 6 mice/group) and compared by one-way ANOVA, followed by the Tukey’s post hoc test. * p < 0.05, ** p < 0.01, versus TBI group; # p < 0.05, sh-TSG-6 hUMSC group versus sh-NC hUMSC group

    Journal: Cell Communication and Signaling : CCS

    Article Title: Mesenchymal stem cells protect against TBI-induced pyroptosis in vivo and in vitro through TSG-6

    doi: 10.1186/s12964-022-00931-2

    Figure Lengend Snippet: hUMSCs downregulated the NLRP3/Caspase-1/GSDMD signaling pathway and reduced microglia pyroptosis in TBI mice via TSG-6. TBI mice were intracerebroventricularly injected with 10 μL of PBS, 0.5 µg rmTSG-6 or 1 µg rmTSG-6, or 2.5 × 10 5 hUMSCs, sh-TSG-6 hUMSCs, sh-NC hUMSCs, in 10 μL of PBS at 4 h after TBI surgery. Then TBI mice were sacrificed and brain tissues were removed at 48 h post-TBI surgery. a Representative protein bands and corresponding grayscale values of NLRP3, pro-Caspase-1, Caspase-1 p20, GSDMD, and cleaved-GSDMD performed by western blotting. b – d Representative double immunofluorescence staining photographs and quantification of Iba-1 (green) with NLRP3 (red), Caspase-1 p20 (red), or GSDMD (red) in the injured cortex. Scale bar = 50 μm. All data are represented as means ± SD (n = 6 mice/group) and compared by one-way ANOVA, followed by the Tukey’s post hoc test. * p < 0.05, ** p < 0.01, versus TBI group; # p < 0.05, sh-TSG-6 hUMSC group versus sh-NC hUMSC group

    Article Snippet: The membranes were blocked for 2 h at 37 °C with 5% non-fat milk and 0.05% Tween-20, then incubated overnight at 4 °C with Mouse anti-GAPDH (60004-1-Ig, Proteintech, 1:5000), rabbit anti-NLRP3 (#15101, CST, 1:1000), rabbit anti-pro-Caspase-1 (AF5418, Affinity, 1:1000), rabbit anti-Caspase-1 p20 (AF4005, Affinity, 1:1000), rabbit anti-GSDMD (ab219800, Abcam, 1:1000), and rabbit anti-cleaved-GSDMD (#10137, CST, 1:1000).

    Techniques: Injection, Western Blot, Double Immunofluorescence Staining

    hUMSCs reduced LPS + ATP-induced BV2 microglia pyroptosis by secreting TSG-6. BV2 cells were co-cultured with hUMSCs, sh-TSG-6 hUMSCs, or sh-NC hUMSCs activated by TNF-α and different concentration of rmTSG-6 (50 ng/mL, 100 ng/mL, 200 ng/mL) and treated with or without LPS (1 µg/mL) + ATP (5 mM) for 12 h. a Representative photomicrographs and quantification of double staining with PI (red) and DAPI (blue) were captured by confocal microscopy. b Caspase-1 activity and the release of LDH in the supernatant was assessed using Caspase-1 activity and LDH activity detection kits. c The protein levels of IL-1β, pro IL-18, and TNF-α were evaluated by ELISA. d Representative protein bands and corresponding grayscale values of NLRP3, pro-Caspase-1, Caspase-1 p20, GSDMD and cleaved-GSDMD were performed by western blotting. Scale bar = 100 μm. All data are represented as means ± SD of at least 3 independent experiments and compared by one-way ANOVA, followed by the Tukey’s post hoc test. * p < 0.05, ** p < 0.01, versus LPS + ATP group; # p < 0.05, sh-TSG-6 hUMSC group versus sh-NC hUMSC group

    Journal: Cell Communication and Signaling : CCS

    Article Title: Mesenchymal stem cells protect against TBI-induced pyroptosis in vivo and in vitro through TSG-6

    doi: 10.1186/s12964-022-00931-2

    Figure Lengend Snippet: hUMSCs reduced LPS + ATP-induced BV2 microglia pyroptosis by secreting TSG-6. BV2 cells were co-cultured with hUMSCs, sh-TSG-6 hUMSCs, or sh-NC hUMSCs activated by TNF-α and different concentration of rmTSG-6 (50 ng/mL, 100 ng/mL, 200 ng/mL) and treated with or without LPS (1 µg/mL) + ATP (5 mM) for 12 h. a Representative photomicrographs and quantification of double staining with PI (red) and DAPI (blue) were captured by confocal microscopy. b Caspase-1 activity and the release of LDH in the supernatant was assessed using Caspase-1 activity and LDH activity detection kits. c The protein levels of IL-1β, pro IL-18, and TNF-α were evaluated by ELISA. d Representative protein bands and corresponding grayscale values of NLRP3, pro-Caspase-1, Caspase-1 p20, GSDMD and cleaved-GSDMD were performed by western blotting. Scale bar = 100 μm. All data are represented as means ± SD of at least 3 independent experiments and compared by one-way ANOVA, followed by the Tukey’s post hoc test. * p < 0.05, ** p < 0.01, versus LPS + ATP group; # p < 0.05, sh-TSG-6 hUMSC group versus sh-NC hUMSC group

    Article Snippet: The membranes were blocked for 2 h at 37 °C with 5% non-fat milk and 0.05% Tween-20, then incubated overnight at 4 °C with Mouse anti-GAPDH (60004-1-Ig, Proteintech, 1:5000), rabbit anti-NLRP3 (#15101, CST, 1:1000), rabbit anti-pro-Caspase-1 (AF5418, Affinity, 1:1000), rabbit anti-Caspase-1 p20 (AF4005, Affinity, 1:1000), rabbit anti-GSDMD (ab219800, Abcam, 1:1000), and rabbit anti-cleaved-GSDMD (#10137, CST, 1:1000).

    Techniques: Cell Culture, Concentration Assay, Double Staining, Confocal Microscopy, Activity Assay, Enzyme-linked Immunosorbent Assay, Western Blot

    Gasdermin d -deficient mice ( Gsdmd -/- ) have reduced pulmonary inflammation and remodeling after acute smoke cigarette (CS) exposure. WT and Gsdmd -/- mice were exposed to CS or Air during 4 days. Total cells (A) , macrophages (B) and neutrophils (C) counts and MPO level in BAL (D) are shown. CXCL1 (E) , CXCL5 (F) , CXCL15 (G) and IL-1β (H) levels were measured in BAL. MMP-9 (I) and TIMP-1 (J) remodeling factor levels were measured in BAL. In lungs, levels of IL-1β (K) , MMP-9 (L) and TIMP-1 (M) were also measured. All parameters decreased in Gsdmd -/- mice exposed to CS as compared to CS WT mice. Data are representative of four experiments and are expressed as mean values ± SEM (n= 4-6 mice per group, *p < 0.05, **p < 0.01, ns, non significant, using a Mann Whitney test).

    Journal: Frontiers in Immunology

    Article Title: Cigarette smoke-induced gasdermin D activation in bronchoalveolar macrophages and bronchial epithelial cells dependently on NLRP3

    doi: 10.3389/fimmu.2022.918507

    Figure Lengend Snippet: Gasdermin d -deficient mice ( Gsdmd -/- ) have reduced pulmonary inflammation and remodeling after acute smoke cigarette (CS) exposure. WT and Gsdmd -/- mice were exposed to CS or Air during 4 days. Total cells (A) , macrophages (B) and neutrophils (C) counts and MPO level in BAL (D) are shown. CXCL1 (E) , CXCL5 (F) , CXCL15 (G) and IL-1β (H) levels were measured in BAL. MMP-9 (I) and TIMP-1 (J) remodeling factor levels were measured in BAL. In lungs, levels of IL-1β (K) , MMP-9 (L) and TIMP-1 (M) were also measured. All parameters decreased in Gsdmd -/- mice exposed to CS as compared to CS WT mice. Data are representative of four experiments and are expressed as mean values ± SEM (n= 4-6 mice per group, *p < 0.05, **p < 0.01, ns, non significant, using a Mann Whitney test).

    Article Snippet: Cells were washed 3 times in TBS, incubated 15 min in TBS-0.3% Triton X-100, then washed 3 times in TBS, blocked in TBS-10% Bovine Serum Albumin (BSA) for 45min and incubated overnight with primary rabbit anti-Cleaved Caspase1 (1:100, #89332, Cell Signaling) or anti-Cleaved Gasdermin (1:100, #10137, Cell Signaling).

    Techniques: MANN-WHITNEY

    Gasdermin d -deficient mice ( Gsdmd -/- ) have reduced pulmonary inflammation and remodeling after acute smoke cigarette (CS) exposure. WT and Gsdmd -/- mice were exposed to CS or Air during 4 days. Total cells (A) , macrophages (B) and neutrophils (C) counts and MPO level in BAL (D) are shown. CXCL1 (E) , CXCL5 (F) , CXCL15 (G) and IL-1β (H) levels were measured in BAL. MMP-9 (I) and TIMP-1 (J) remodeling factor levels were measured in BAL. In lungs, levels of IL-1β (K) , MMP-9 (L) and TIMP-1 (M) were also measured. All parameters decreased in Gsdmd -/- mice exposed to CS as compared to CS WT mice. Data are representative of four experiments and are expressed as mean values ± SEM (n= 4-6 mice per group, *p < 0.05, **p < 0.01, ns, non significant, using a Mann Whitney test).

    Journal: Frontiers in Immunology

    Article Title: Cigarette smoke-induced gasdermin D activation in bronchoalveolar macrophages and bronchial epithelial cells dependently on NLRP3

    doi: 10.3389/fimmu.2022.918507

    Figure Lengend Snippet: Gasdermin d -deficient mice ( Gsdmd -/- ) have reduced pulmonary inflammation and remodeling after acute smoke cigarette (CS) exposure. WT and Gsdmd -/- mice were exposed to CS or Air during 4 days. Total cells (A) , macrophages (B) and neutrophils (C) counts and MPO level in BAL (D) are shown. CXCL1 (E) , CXCL5 (F) , CXCL15 (G) and IL-1β (H) levels were measured in BAL. MMP-9 (I) and TIMP-1 (J) remodeling factor levels were measured in BAL. In lungs, levels of IL-1β (K) , MMP-9 (L) and TIMP-1 (M) were also measured. All parameters decreased in Gsdmd -/- mice exposed to CS as compared to CS WT mice. Data are representative of four experiments and are expressed as mean values ± SEM (n= 4-6 mice per group, *p < 0.05, **p < 0.01, ns, non significant, using a Mann Whitney test).

    Article Snippet: Lung sections were permeabilized in PBS 0.5% triton X-100, blocked with 5% FCS for 1h at RT and then incubated overnight with primary mouse anti-cleaved Gasdermin D (1:100, #10137, Cell Signaling).

    Techniques: MANN-WHITNEY