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antibodies against gsdmd n  (Affinity Biosciences)

 
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    Affinity Biosciences antibodies against gsdmd n
    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, <t>GSDMD-N,</t> 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.
    Antibodies Against Gsdmd N, 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/gsdmd+n/anti+gsdmd+n/pmc12907102-318-0-19
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    antibodies against gsdmd n - by Bioz Stars, 2026-10
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    Images

    1) Product Images from "3D-MSCs apoptotic bodies-integrated conductive hydrogel mitigates spinal cord injury via immunoregulation and alleviating neuronal pyroptosis"

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

    Journal: Bioactive Materials

    doi: 10.1016/j.bioactmat.2026.01.043

    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.
    Figure Legend 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.

    Techniques Used: 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.
    Figure Legend 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.

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

    Related Articles

    Blocking Assay:

    Article Title: Inhibition of ASIC1a Attenuates Neuronal Pyroptosis and Neuroinflammation Following Traumatic Brain Injury.
    Article Snippet: .. Sections were blocked with QuickBlock Blocking Buffer for Immunol Staining (Beyotime, P0260, China) at room temperature for 10 min, and incubated overnight at 4°C with primary antibodies: ASIC1a (1:100; Proteintech, 27235- 1- AP), NeuN (1:100, Proteintech, 26975- 1- AP), Iba1 (1:200, Proteintech, 10904- 1- AP), GFAP (1:200, Proteintech, 16825- 1- AP), GSDMD- N (1:100; Affinity Biosciences, DF13758). .. The following day, sections were rewarmed at room temperature for 30 min, then washed with phosphate- buffered saline (PBS) three times, and incubated with secondary antibody (Alexa Fluor 594 and Alexa Fluor 488, Abcam, ab150080 and ab 150077) at 37°C for 1 h. Then, sections were washed with PBS three times, stained with DAPI for 15 min, and then washed with PBS.

    Article Title: Inhibition of ASIC1a Attenuates Neuronal Pyroptosis and Neuroinflammation Following Traumatic Brain Injury
    Article Snippet: .. Sections were blocked with QuickBlock Blocking Buffer for Immunol Staining (Beyotime, P0260, China) at room temperature for 10 min, and incubated overnight at 4°C with primary antibodies: ASIC1a (1:100; Proteintech, 27235‐1‐AP), NeuN (1:100, Proteintech, 26975‐1‐AP), Iba1 (1:200, Proteintech, 10904‐1‐AP), GFAP (1:200, Proteintech, 16825‐1‐AP), GSDMD‐N (1:100; Affinity Biosciences, DF13758). .. The following day, sections were rewarmed at room temperature for 30 min, then washed with phosphate‐buffered saline (PBS) three times, and incubated with secondary antibody (Alexa Fluor 594 and Alexa Fluor 488, Abcam, ab150080 and ab 150077) at 37°C for 1 h. Then, sections were washed with PBS three times, stained with DAPI for 15 min, and then washed with PBS.

    Staining:

    Article Title: Inhibition of ASIC1a Attenuates Neuronal Pyroptosis and Neuroinflammation Following Traumatic Brain Injury.
    Article Snippet: .. Sections were blocked with QuickBlock Blocking Buffer for Immunol Staining (Beyotime, P0260, China) at room temperature for 10 min, and incubated overnight at 4°C with primary antibodies: ASIC1a (1:100; Proteintech, 27235- 1- AP), NeuN (1:100, Proteintech, 26975- 1- AP), Iba1 (1:200, Proteintech, 10904- 1- AP), GFAP (1:200, Proteintech, 16825- 1- AP), GSDMD- N (1:100; Affinity Biosciences, DF13758). .. The following day, sections were rewarmed at room temperature for 30 min, then washed with phosphate- buffered saline (PBS) three times, and incubated with secondary antibody (Alexa Fluor 594 and Alexa Fluor 488, Abcam, ab150080 and ab 150077) at 37°C for 1 h. Then, sections were washed with PBS three times, stained with DAPI for 15 min, and then washed with PBS.

    Article Title: Inhibition of ASIC1a Attenuates Neuronal Pyroptosis and Neuroinflammation Following Traumatic Brain Injury
    Article Snippet: .. Sections were blocked with QuickBlock Blocking Buffer for Immunol Staining (Beyotime, P0260, China) at room temperature for 10 min, and incubated overnight at 4°C with primary antibodies: ASIC1a (1:100; Proteintech, 27235‐1‐AP), NeuN (1:100, Proteintech, 26975‐1‐AP), Iba1 (1:200, Proteintech, 10904‐1‐AP), GFAP (1:200, Proteintech, 16825‐1‐AP), GSDMD‐N (1:100; Affinity Biosciences, DF13758). .. The following day, sections were rewarmed at room temperature for 30 min, then washed with phosphate‐buffered saline (PBS) three times, and incubated with secondary antibody (Alexa Fluor 594 and Alexa Fluor 488, Abcam, ab150080 and ab 150077) at 37°C for 1 h. Then, sections were washed with PBS three times, stained with DAPI for 15 min, and then washed with PBS.

    Incubation:

    Article Title: Inhibition of ASIC1a Attenuates Neuronal Pyroptosis and Neuroinflammation Following Traumatic Brain Injury.
    Article Snippet: .. Sections were blocked with QuickBlock Blocking Buffer for Immunol Staining (Beyotime, P0260, China) at room temperature for 10 min, and incubated overnight at 4°C with primary antibodies: ASIC1a (1:100; Proteintech, 27235- 1- AP), NeuN (1:100, Proteintech, 26975- 1- AP), Iba1 (1:200, Proteintech, 10904- 1- AP), GFAP (1:200, Proteintech, 16825- 1- AP), GSDMD- N (1:100; Affinity Biosciences, DF13758). .. The following day, sections were rewarmed at room temperature for 30 min, then washed with phosphate- buffered saline (PBS) three times, and incubated with secondary antibody (Alexa Fluor 594 and Alexa Fluor 488, Abcam, ab150080 and ab 150077) at 37°C for 1 h. Then, sections were washed with PBS three times, stained with DAPI for 15 min, and then washed with PBS.

    Article Title: Melatonin protects human nucleus pulposus cells from pyroptosis by regulating Nrf2 via melatonin membrane receptors
    Article Snippet: .. After transfer, the membranes were blocked in Tris- buffered saline (TBST) containing 5% non- fat milk for two hours at 37°C and then incubated overnight with primary anti- NLRP3 (1:1,000; Abcam, USA, Cat. No.: ab210491), CASP- 1 (1:1,000, Proteintech, USA, Cat. No.: 22915- 1- AP), GSDMD (1:1,000, Affinity Biosciences, USA, Cat. #: AF4012), GSDMD- N (1:1,000, Affinity Biosciences, Cat. #: DF12275), IL- 18 (1:1,000, Affinity Biosciences, Cat. #: DF6252), IL- 1β (1:500, ABclonal, USA, Cat. No.: A1112), Nrf2 (1:1,000, Abcam, Cat. No.: ab62352), and glyceraldehyde 3- phosphate dehydrogenase (GAPDH) (1:10,000, Proteintech, Cat. No.: 10494- 1- AP) at 4°C. ..

    Article Title: Inhibition of ASIC1a Attenuates Neuronal Pyroptosis and Neuroinflammation Following Traumatic Brain Injury
    Article Snippet: .. Sections were blocked with QuickBlock Blocking Buffer for Immunol Staining (Beyotime, P0260, China) at room temperature for 10 min, and incubated overnight at 4°C with primary antibodies: ASIC1a (1:100; Proteintech, 27235‐1‐AP), NeuN (1:100, Proteintech, 26975‐1‐AP), Iba1 (1:200, Proteintech, 10904‐1‐AP), GFAP (1:200, Proteintech, 16825‐1‐AP), GSDMD‐N (1:100; Affinity Biosciences, DF13758). .. The following day, sections were rewarmed at room temperature for 30 min, then washed with phosphate‐buffered saline (PBS) three times, and incubated with secondary antibody (Alexa Fluor 594 and Alexa Fluor 488, Abcam, ab150080 and ab 150077) at 37°C for 1 h. Then, sections were washed with PBS three times, stained with DAPI for 15 min, and then washed with PBS.

    Article Title: Dietary Zinc activates the Nrf2 signaling pathway to inhibit pyroptosis and attenuate the lung inflammatory response in COPD.
    Article Snippet: Pyroptosis and inflammation play crucial roles in the development of chronic obstructive pulmonary disease (COPD), and Zinc deficiency is commonly observed in COPD patients.. In this study, we aimed to explore the impact of Zinc supplementation on pyroptosis and inflammation in a cigarette smoke (CS)-induced COPD mouse model, as well as the underlying mechanisms.. The COPD mouse model was established through CS exposure, and mouse pulmonary epithelial cells (MLE-12) were exposed to cigarette smoke extract (CSE) to further validate the effects of Zinc supplementation.

    Article Title: Knockdown of PHLDA1 Alleviates Necrotizing Enterocolitis by Inhibiting NLRP3 Inflammasome Activation and Pyroptosis Through Enhancing Nrf2 Signaling.
    Article Snippet: Objective: Pleckstrin homology-like domain family A member 1 (PHLDA1) is involved in the progression of intestine-related diseases, but its role and related mechanisms in Necrotizing enterocolitis (NEC) are unclear.. The aim of this study was to better understand the function of PHLDA1 in NEC and the underlying mechanisms.. Methods: A neonatal mouse model of NEC was established by hypoxic hypothermia, and sh-PHLDA1 was transfected into mice to observe the mortality of each group within 4 days.

    Saline:

    Article Title: Melatonin protects human nucleus pulposus cells from pyroptosis by regulating Nrf2 via melatonin membrane receptors
    Article Snippet: .. After transfer, the membranes were blocked in Tris- buffered saline (TBST) containing 5% non- fat milk for two hours at 37°C and then incubated overnight with primary anti- NLRP3 (1:1,000; Abcam, USA, Cat. No.: ab210491), CASP- 1 (1:1,000, Proteintech, USA, Cat. No.: 22915- 1- AP), GSDMD (1:1,000, Affinity Biosciences, USA, Cat. #: AF4012), GSDMD- N (1:1,000, Affinity Biosciences, Cat. #: DF12275), IL- 18 (1:1,000, Affinity Biosciences, Cat. #: DF6252), IL- 1β (1:500, ABclonal, USA, Cat. No.: A1112), Nrf2 (1:1,000, Abcam, Cat. No.: ab62352), and glyceraldehyde 3- phosphate dehydrogenase (GAPDH) (1:10,000, Proteintech, Cat. No.: 10494- 1- AP) at 4°C. ..

    Membrane:

    Article Title: Knockdown of PHLDA1 Alleviates Necrotizing Enterocolitis by Inhibiting NLRP3 Inflammasome Activation and Pyroptosis Through Enhancing Nrf2 Signaling.
    Article Snippet: Objective: Pleckstrin homology-like domain family A member 1 (PHLDA1) is involved in the progression of intestine-related diseases, but its role and related mechanisms in Necrotizing enterocolitis (NEC) are unclear.. The aim of this study was to better understand the function of PHLDA1 in NEC and the underlying mechanisms.. Methods: A neonatal mouse model of NEC was established by hypoxic hypothermia, and sh-PHLDA1 was transfected into mice to observe the mortality of each group within 4 days.

    Expressing:

    Article Title: Knockdown of PHLDA1 Alleviates Necrotizing Enterocolitis by Inhibiting NLRP3 Inflammasome Activation and Pyroptosis Through Enhancing Nrf2 Signaling.
    Article Snippet: Objective: Pleckstrin homology-like domain family A member 1 (PHLDA1) is involved in the progression of intestine-related diseases, but its role and related mechanisms in Necrotizing enterocolitis (NEC) are unclear.. The aim of this study was to better understand the function of PHLDA1 in NEC and the underlying mechanisms.. Methods: A neonatal mouse model of NEC was established by hypoxic hypothermia, and sh-PHLDA1 was transfected into mice to observe the mortality of each group within 4 days.

    Control:

    Article Title: Knockdown of PHLDA1 Alleviates Necrotizing Enterocolitis by Inhibiting NLRP3 Inflammasome Activation and Pyroptosis Through Enhancing Nrf2 Signaling.
    Article Snippet: Objective: Pleckstrin homology-like domain family A member 1 (PHLDA1) is involved in the progression of intestine-related diseases, but its role and related mechanisms in Necrotizing enterocolitis (NEC) are unclear.. The aim of this study was to better understand the function of PHLDA1 in NEC and the underlying mechanisms.. Methods: A neonatal mouse model of NEC was established by hypoxic hypothermia, and sh-PHLDA1 was transfected into mice to observe the mortality of each group within 4 days.



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    94
    MedChemExpress gsdmd n
    In vivo therapeutic efficacy of MPB@TA‐Cu‐Ma in an intestinal I/R injury model and the mechanism of GSDMD‐mediated pyroptosis. (A) Schematic illustration of the experimental timeline for the intestinal ischemia (45 min) and reperfusion (6 h) model in C57BL/6 mice. (B) Representative H&E‐stained images and histopathological Chiu scores of intestinal tissues from different groups. Scale bar = 200 µm. (C) ROS fluorescence signal (green) in intestinal tissues detected using a fluorescent probe (DCFH‐DA), and quantitative histograms; scale bars: 200 µm. (D, E) Relative SOD activity and MDA concentration in intestinal tissues ( n = 5 biologically independent mice per group). (F–I) Serum concentrations of pro‐inflammatory cytokines TNF‐α (F), IL‐6 (G), IL‐1β (H), and IL‐18 (I) across different experimental groups as measured by ELISA. (J) Representative immunohistochemistry (IHC) images and quantitative optical density analysis <t>of</t> <t>GSDMD‐N</t> expression in intestinal tissues. Scale bar = 200 µm. (K) HE‐stained small intestinal sections from WT and Gsdmd ΔMΦ mice in Control and I/R groups, with quantitative injury analysis using the Chiu score. Scale bar: 200 µm. (L) ROS fluorescence in frozen small intestinal sections from WT, Gsdmd ΔMΦ , and Gsdmd ΔMΦ + MPB@TA‐Cu‐Ma mice after I/R injury, with quantitative analysis of fluorescence intensity. Scale bar: 100 µm. (M‐N) Quantitative analysis of serum IL‐6 and TNF‐α levels in WT and Gsdmd ΔMΦ C57BL/6 mice from Control and I/R groups. Data are presented as mean ± SEM from biologically independent mice ( n = 4 for B, K; n = 5 for D–E; n = 3 for all other panels). Statistical analysis was performed using one‐way ANOVA (C–J, L) and two‐way ANOVA (M–N) followed by Tukey's post hoc test. For ordinal data (Chiu scores in B and K), statistical significance was determined using the non‐parametric Kruskal–Wallis test followed by Dunn's multiple comparisons test (B) and the Mann–Whitney U test (K). **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05; ns, not significant.
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    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: Antibodies against GSDMD-N (cat. no. DF12275), CD44 (cat. no. BF9213), and Integrin beta1 (cat. no. AF5379) were obtained from Affinity Biosciences in Ohio, 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: Antibodies against GSDMD-N (cat. no. DF12275), CD44 (cat. no. BF9213), and Integrin beta1 (cat. no. AF5379) were obtained from Affinity Biosciences in Ohio, USA.

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

    MaR1 ameliorates HG-induced pyroptosis in ARPE-19 cells. (A) Effect of MaR1 treatment at varying concentrations on the fluorescence intensity of GSDMD-N in HG-induced ARPE-19 cells. Scale bar, 50 µm. (B) Effect of MaR1 treatment at varying concentrations on the expression of pyroptosis-related proteins (caspase-1, GSDMD, GSDMD-N, cleaved caspase-1, NLRP3, ASC and IL-18) in HG-induced ARPE-19 cells. * P<0.05, ** P<0.01 and *** P<0.001. MaR1, maresin 1; HG, high glucose; NG, normal glucose; GSDMD, gasdermin D; GSDMD-N, GSDMD N-terminal; NLRP3, NLR family pyrin domain containing 3; ASC, apoptosis-associated speck-like protein containing a CARD.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Maresin 1 activates autophagy through SIRT1/PPAR-γ signaling to mitigate high glucose-induced pyroptosis in human retinal pigment epithelial cells

    doi: 10.3892/etm.2026.13134

    Figure Lengend Snippet: MaR1 ameliorates HG-induced pyroptosis in ARPE-19 cells. (A) Effect of MaR1 treatment at varying concentrations on the fluorescence intensity of GSDMD-N in HG-induced ARPE-19 cells. Scale bar, 50 µm. (B) Effect of MaR1 treatment at varying concentrations on the expression of pyroptosis-related proteins (caspase-1, GSDMD, GSDMD-N, cleaved caspase-1, NLRP3, ASC and IL-18) in HG-induced ARPE-19 cells. * P<0.05, ** P<0.01 and *** P<0.001. MaR1, maresin 1; HG, high glucose; NG, normal glucose; GSDMD, gasdermin D; GSDMD-N, GSDMD N-terminal; NLRP3, NLR family pyrin domain containing 3; ASC, apoptosis-associated speck-like protein containing a CARD.

    Article Snippet: Subsequently, the cells were blocked with 1% BSA solution (cat. no. ST023; Beyotime Biotechnology) at room temperature for 1 h, followed by overnight incubation at 4 ̊C with diluted anti-GSDMD N-terminal (GSDMD-N) primary antibody (1:100; cat. no. ER1901-37; HUABIO).

    Techniques: Fluorescence, Expressing

    MaR1 mediates autophagy through SIRT1/peroxisome proliferator-activated receptor-γ signaling to ameliorate HG-induced pyroptosis in ARPE-19 cells. (A) SIRT1 inhibitor EX527 and autophagy inhibitor 3-MA enhanced the fluorescence intensity of GSDMD-N in ARPE-19 cells treated with HG and MaR1. Scale bar, 50 µm. (B) EX527 and 3-MA upregulated the expression of GSDMD-N, cleaved caspase-1, NLRP3, ASC and IL-18 in ARPE-19 cells treated with HG and MaR1. * P<0.05, ** P<0.01 and *** P<0.001. MaR1, maresin 1; HG, high glucose; NG, normal glucose; SIRT1, sirtuin 1; GSDMD, gasdermin D; GSDMD-N, GSDMD N-terminal; NLRP3, NLR family pyrin domain containing 3; ASC, apoptosis-associated speck-like protein containing a CARD; 3-MA, 3-methyladenine.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Maresin 1 activates autophagy through SIRT1/PPAR-γ signaling to mitigate high glucose-induced pyroptosis in human retinal pigment epithelial cells

    doi: 10.3892/etm.2026.13134

    Figure Lengend Snippet: MaR1 mediates autophagy through SIRT1/peroxisome proliferator-activated receptor-γ signaling to ameliorate HG-induced pyroptosis in ARPE-19 cells. (A) SIRT1 inhibitor EX527 and autophagy inhibitor 3-MA enhanced the fluorescence intensity of GSDMD-N in ARPE-19 cells treated with HG and MaR1. Scale bar, 50 µm. (B) EX527 and 3-MA upregulated the expression of GSDMD-N, cleaved caspase-1, NLRP3, ASC and IL-18 in ARPE-19 cells treated with HG and MaR1. * P<0.05, ** P<0.01 and *** P<0.001. MaR1, maresin 1; HG, high glucose; NG, normal glucose; SIRT1, sirtuin 1; GSDMD, gasdermin D; GSDMD-N, GSDMD N-terminal; NLRP3, NLR family pyrin domain containing 3; ASC, apoptosis-associated speck-like protein containing a CARD; 3-MA, 3-methyladenine.

    Article Snippet: Subsequently, the cells were blocked with 1% BSA solution (cat. no. ST023; Beyotime Biotechnology) at room temperature for 1 h, followed by overnight incubation at 4 ̊C with diluted anti-GSDMD N-terminal (GSDMD-N) primary antibody (1:100; cat. no. ER1901-37; HUABIO).

    Techniques: Fluorescence, Expressing

    Schematic illustration of the protective mechanism of MaR1 against HG-induced pyroptosis in ARPE-19 cells. High glucose stimulation triggers the overproduction of ROS, leading to the activation of the NLRP3 inflammasome complex (comprising NLRP3, ASC and Caspase-1). This activation results in the cleavage of GSDMD, which forms membrane pores (GSDMD-N) and promotes the release of pro-inflammatory cytokines, ultimately inducing pyroptosis. However, MaR1 treatment effectively upregulates SIRT1 expression, which in turn enhances PPAR-γ levels and restores autophagic flux. The restored autophagy facilitates the clearance of damaged mitochondria and ROS, thereby inhibiting the NLRP3/GSDMD signaling pathway and protecting ARPE-19 cells from HG-induced inflammatory damage. SIRT1, sirtuin 1; PPAR-γ, peroxisome proliferator-activated receptor-γ; ROS, reactive oxygen species; GSDMD-N, gasdermin D N-terminal; NLRP3, NLR family pyrin domain containing 3; ASC, apoptosis-associated speck-like protein containing a CARD.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Maresin 1 activates autophagy through SIRT1/PPAR-γ signaling to mitigate high glucose-induced pyroptosis in human retinal pigment epithelial cells

    doi: 10.3892/etm.2026.13134

    Figure Lengend Snippet: Schematic illustration of the protective mechanism of MaR1 against HG-induced pyroptosis in ARPE-19 cells. High glucose stimulation triggers the overproduction of ROS, leading to the activation of the NLRP3 inflammasome complex (comprising NLRP3, ASC and Caspase-1). This activation results in the cleavage of GSDMD, which forms membrane pores (GSDMD-N) and promotes the release of pro-inflammatory cytokines, ultimately inducing pyroptosis. However, MaR1 treatment effectively upregulates SIRT1 expression, which in turn enhances PPAR-γ levels and restores autophagic flux. The restored autophagy facilitates the clearance of damaged mitochondria and ROS, thereby inhibiting the NLRP3/GSDMD signaling pathway and protecting ARPE-19 cells from HG-induced inflammatory damage. SIRT1, sirtuin 1; PPAR-γ, peroxisome proliferator-activated receptor-γ; ROS, reactive oxygen species; GSDMD-N, gasdermin D N-terminal; NLRP3, NLR family pyrin domain containing 3; ASC, apoptosis-associated speck-like protein containing a CARD.

    Article Snippet: Subsequently, the cells were blocked with 1% BSA solution (cat. no. ST023; Beyotime Biotechnology) at room temperature for 1 h, followed by overnight incubation at 4 ̊C with diluted anti-GSDMD N-terminal (GSDMD-N) primary antibody (1:100; cat. no. ER1901-37; HUABIO).

    Techniques: Activation Assay, Membrane, Expressing

    In vivo therapeutic efficacy of MPB@TA‐Cu‐Ma in an intestinal I/R injury model and the mechanism of GSDMD‐mediated pyroptosis. (A) Schematic illustration of the experimental timeline for the intestinal ischemia (45 min) and reperfusion (6 h) model in C57BL/6 mice. (B) Representative H&E‐stained images and histopathological Chiu scores of intestinal tissues from different groups. Scale bar = 200 µm. (C) ROS fluorescence signal (green) in intestinal tissues detected using a fluorescent probe (DCFH‐DA), and quantitative histograms; scale bars: 200 µm. (D, E) Relative SOD activity and MDA concentration in intestinal tissues ( n = 5 biologically independent mice per group). (F–I) Serum concentrations of pro‐inflammatory cytokines TNF‐α (F), IL‐6 (G), IL‐1β (H), and IL‐18 (I) across different experimental groups as measured by ELISA. (J) Representative immunohistochemistry (IHC) images and quantitative optical density analysis of GSDMD‐N expression in intestinal tissues. Scale bar = 200 µm. (K) HE‐stained small intestinal sections from WT and Gsdmd ΔMΦ mice in Control and I/R groups, with quantitative injury analysis using the Chiu score. Scale bar: 200 µm. (L) ROS fluorescence in frozen small intestinal sections from WT, Gsdmd ΔMΦ , and Gsdmd ΔMΦ + MPB@TA‐Cu‐Ma mice after I/R injury, with quantitative analysis of fluorescence intensity. Scale bar: 100 µm. (M‐N) Quantitative analysis of serum IL‐6 and TNF‐α levels in WT and Gsdmd ΔMΦ C57BL/6 mice from Control and I/R groups. Data are presented as mean ± SEM from biologically independent mice ( n = 4 for B, K; n = 5 for D–E; n = 3 for all other panels). Statistical analysis was performed using one‐way ANOVA (C–J, L) and two‐way ANOVA (M–N) followed by Tukey's post hoc test. For ordinal data (Chiu scores in B and K), statistical significance was determined using the non‐parametric Kruskal–Wallis test followed by Dunn's multiple comparisons test (B) and the Mann–Whitney U test (K). **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05; ns, not significant.

    Journal: Advanced Science

    Article Title: Tailored Porous Bimetallic Nanozyme Platform for Full‐Cycle Therapeutics of Intestinal Ischemia/Reperfusion

    doi: 10.1002/advs.202520748

    Figure Lengend Snippet: In vivo therapeutic efficacy of MPB@TA‐Cu‐Ma in an intestinal I/R injury model and the mechanism of GSDMD‐mediated pyroptosis. (A) Schematic illustration of the experimental timeline for the intestinal ischemia (45 min) and reperfusion (6 h) model in C57BL/6 mice. (B) Representative H&E‐stained images and histopathological Chiu scores of intestinal tissues from different groups. Scale bar = 200 µm. (C) ROS fluorescence signal (green) in intestinal tissues detected using a fluorescent probe (DCFH‐DA), and quantitative histograms; scale bars: 200 µm. (D, E) Relative SOD activity and MDA concentration in intestinal tissues ( n = 5 biologically independent mice per group). (F–I) Serum concentrations of pro‐inflammatory cytokines TNF‐α (F), IL‐6 (G), IL‐1β (H), and IL‐18 (I) across different experimental groups as measured by ELISA. (J) Representative immunohistochemistry (IHC) images and quantitative optical density analysis of GSDMD‐N expression in intestinal tissues. Scale bar = 200 µm. (K) HE‐stained small intestinal sections from WT and Gsdmd ΔMΦ mice in Control and I/R groups, with quantitative injury analysis using the Chiu score. Scale bar: 200 µm. (L) ROS fluorescence in frozen small intestinal sections from WT, Gsdmd ΔMΦ , and Gsdmd ΔMΦ + MPB@TA‐Cu‐Ma mice after I/R injury, with quantitative analysis of fluorescence intensity. Scale bar: 100 µm. (M‐N) Quantitative analysis of serum IL‐6 and TNF‐α levels in WT and Gsdmd ΔMΦ C57BL/6 mice from Control and I/R groups. Data are presented as mean ± SEM from biologically independent mice ( n = 4 for B, K; n = 5 for D–E; n = 3 for all other panels). Statistical analysis was performed using one‐way ANOVA (C–J, L) and two‐way ANOVA (M–N) followed by Tukey's post hoc test. For ordinal data (Chiu scores in B and K), statistical significance was determined using the non‐parametric Kruskal–Wallis test followed by Dunn's multiple comparisons test (B) and the Mann–Whitney U test (K). **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05; ns, not significant.

    Article Snippet: The sections were then incubated with primary antibodies against TNF‐α (Proteintech, 26405‐1‐AP, 1:200, RRID:AB_2918102), GSDMD‐N (Immunoway, YT7991, 1:100, RRID:AB_3663000), Caspase‐1 (Biodragon, BD‐PT5743, 1:200), NLPR3 (MCE, HY‐ P80246 , 1:200), GSTM1 (Proteintech, 12412‐1‐AP, 1:200, RRID:AB_2115925), NOX1 (Biodragon, BD‐PT2820, 1:200), VEGF (Proteintech, 19003‐1‐AP, 1:200, RRID:AB_2212657), ACE (Proteintech, 24743‐1‐AP, 1:200, RRID:AB_2879701), DSG1 ( Biodragon, BD‐PT5886,1:200) and DEFA1 (Biodragon, BD‐PT1322, 1:200) overnight at 4°C, and followed by incubation with biotin‐conjugated anti‐rabbit antibody or anti‐goat antibody.

    Techniques: In Vivo, Drug discovery, Staining, Fluorescence, Activity Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Expressing, Control, MANN-WHITNEY