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rabbit anti nucleotide binding oligomerization domain like receptor protein 3 nlrp3 antibody  (Bioss)


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    Bioss rabbit anti nucleotide binding oligomerization domain like receptor protein 3 nlrp3 antibody
    Rabbit Anti Nucleotide Binding Oligomerization Domain Like Receptor Protein 3 Nlrp3 Antibody, supplied by Bioss, used in various techniques. Bioz Stars score: 95/100, based on 100 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti nucleotide binding oligomerization domain like receptor protein 3 nlrp3 antibody/product/Bioss
    Average 95 stars, based on 100 article reviews
    rabbit anti nucleotide binding oligomerization domain like receptor protein 3 nlrp3 antibody - by Bioz Stars, 2026-03
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    rabbit anti nucleotide binding oligomerization domain like receptor protein 3 nlrp3 antibody  (Bioss)
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    anti nlrp3 antibody  (Bioss)
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    Effect of RASD1 expression on <t>NLRP3</t> mRNA levels in LPS-induced RAW264.7 macrophages. (A) Changes in NLRP3 mRNA expression levels following RASD1 overexpression in Raw264.7 cells. (B) Changes in NLRP3 mRNA expression levels at different RASD1 knockdown sites in Raw264.7 cells. (siRASD1 is the RASD1-knockdown group, and miRASD1 is the RASD1-overexpression group) (Data are presented as mean ± SD from three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns indicates no significant difference.)
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    Effect of nCGA in decreasing both the pyroptosis and inflammation levels in glucolipotoxicity-induced MES-13 cells. (A) MES-13 cells were subjected to Western blotting to analyze <t>NLRP3,</t> GSDMD, IL-1β, and Caspase-1 expression. (B) Quantification of the protein expression. (C) Annexin-V/PI flow cytometry analysis was performed to detect phosphatidylserine externalization, a marker of both apoptosis and pyroptosis-associated membrane disruption. (D–F) Immunofluorescence staining for the expression and cellular localization of pyroptosis markers. Data are shown as mean ± SD. Results were statistically analyzed using ANOVA, followed by the Bonferroni post-hoc test. * p < 0.05 was considered statistically significant compared to the HG + OA group.
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    Effect of RASD1 expression on NLRP3 mRNA levels in LPS-induced RAW264.7 macrophages. (A) Changes in NLRP3 mRNA expression levels following RASD1 overexpression in Raw264.7 cells. (B) Changes in NLRP3 mRNA expression levels at different RASD1 knockdown sites in Raw264.7 cells. (siRASD1 is the RASD1-knockdown group, and miRASD1 is the RASD1-overexpression group) (Data are presented as mean ± SD from three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns indicates no significant difference.)

    Journal: Scientific Reports

    Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

    doi: 10.1038/s41598-025-24410-z

    Figure Lengend Snippet: Effect of RASD1 expression on NLRP3 mRNA levels in LPS-induced RAW264.7 macrophages. (A) Changes in NLRP3 mRNA expression levels following RASD1 overexpression in Raw264.7 cells. (B) Changes in NLRP3 mRNA expression levels at different RASD1 knockdown sites in Raw264.7 cells. (siRASD1 is the RASD1-knockdown group, and miRASD1 is the RASD1-overexpression group) (Data are presented as mean ± SD from three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns indicates no significant difference.)

    Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

    Techniques: Expressing, Over Expression, Knockdown

    Effect of RASD1 expression on NLRP3 inflammasome components and downstream inflammatory factors in LPS-induced RAW264.7 macrophages. (A) mRNA expression of NLRP3 in Raw264.7 cells following RASD1 knockdown and overexpression. (B) mRNA expression of ASC in Raw264.7 cells following RASD1 knockdown and overexpression. (C) mRNA expression of caspase-1 in Raw264.7 cells following RASD1 knockdown and overexpression. (D) mRNA expression of GSDMD in Raw264.7 cells following RASD1 knockdown and overexpression. (E) mRNA expression of IL-1β in Raw264.7 cells following RASD1 knockdown and overexpression. (F) mRNA expression of IL-18 in Raw264.7 cells following RASD1 knockdown and overexpression. Data are presented as mean ± SD from three independent experiments. (siRASD1 is the RASD1 knockdown group, miRASD1 represents the RASD1 overexpression group, * p < 0.05VS.CON; ♆ p < 0.05VS.Model; ◑ p < 0.05VS.HHQG; ▲ p < 0.05VS.Model siRASD1; ● p < 0.05VS.Model miRASD1).

    Journal: Scientific Reports

    Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

    doi: 10.1038/s41598-025-24410-z

    Figure Lengend Snippet: Effect of RASD1 expression on NLRP3 inflammasome components and downstream inflammatory factors in LPS-induced RAW264.7 macrophages. (A) mRNA expression of NLRP3 in Raw264.7 cells following RASD1 knockdown and overexpression. (B) mRNA expression of ASC in Raw264.7 cells following RASD1 knockdown and overexpression. (C) mRNA expression of caspase-1 in Raw264.7 cells following RASD1 knockdown and overexpression. (D) mRNA expression of GSDMD in Raw264.7 cells following RASD1 knockdown and overexpression. (E) mRNA expression of IL-1β in Raw264.7 cells following RASD1 knockdown and overexpression. (F) mRNA expression of IL-18 in Raw264.7 cells following RASD1 knockdown and overexpression. Data are presented as mean ± SD from three independent experiments. (siRASD1 is the RASD1 knockdown group, miRASD1 represents the RASD1 overexpression group, * p < 0.05VS.CON; ♆ p < 0.05VS.Model; ◑ p < 0.05VS.HHQG; ▲ p < 0.05VS.Model siRASD1; ● p < 0.05VS.Model miRASD1).

    Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

    Techniques: Expressing, Knockdown, Over Expression

    Protein expression of NLRP3, RASD1, and GSDMD in LPS-induced RAW264.7 macrophages following RASD1 modulation. (A) Protein expression of NLRP3, RASD1, and GSDMD in Raw264.7 cells following RASD1 overexpression and the results of the statistical analysis of the strip gray value. (B) Protein expression of NLRP3, RASD1, and GSDMD in Raw264.7 cells following RASD1 knockdown and the results of the statistical analysis of the strip gray value. (Each band in the figure represents a different blot, all samples are from the same experiment, and the blots were processed in parallel.) (siRASD1 is the RASD1-knockdown group, and miRASD1 is the RASD1-overexpression group).

    Journal: Scientific Reports

    Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

    doi: 10.1038/s41598-025-24410-z

    Figure Lengend Snippet: Protein expression of NLRP3, RASD1, and GSDMD in LPS-induced RAW264.7 macrophages following RASD1 modulation. (A) Protein expression of NLRP3, RASD1, and GSDMD in Raw264.7 cells following RASD1 overexpression and the results of the statistical analysis of the strip gray value. (B) Protein expression of NLRP3, RASD1, and GSDMD in Raw264.7 cells following RASD1 knockdown and the results of the statistical analysis of the strip gray value. (Each band in the figure represents a different blot, all samples are from the same experiment, and the blots were processed in parallel.) (siRASD1 is the RASD1-knockdown group, and miRASD1 is the RASD1-overexpression group).

    Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

    Techniques: Expressing, Over Expression, Stripping Membranes, Knockdown

    Effect of HHQG on mRNA expression of NLRP3 inflammasome components and downstream inflammatory factors in CCl4-induced liver injury. (A) mRNA expression of NLRP3 in liver tissue. (B) mRNA expression of ASC in liver tissue. (C) mRNA expression of caspase-1 in liver tissue. (D) mRNA expression of IL-18 in liver tissue. (E) mRNA expression of IL-1β in liver tissue. (F) mRNA expression of GSDMD in liver tissue. (G) Correlation analysis between RASD1 and NLRP3 (r = 0.5727, p < 0.05).

    Journal: Scientific Reports

    Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

    doi: 10.1038/s41598-025-24410-z

    Figure Lengend Snippet: Effect of HHQG on mRNA expression of NLRP3 inflammasome components and downstream inflammatory factors in CCl4-induced liver injury. (A) mRNA expression of NLRP3 in liver tissue. (B) mRNA expression of ASC in liver tissue. (C) mRNA expression of caspase-1 in liver tissue. (D) mRNA expression of IL-18 in liver tissue. (E) mRNA expression of IL-1β in liver tissue. (F) mRNA expression of GSDMD in liver tissue. (G) Correlation analysis between RASD1 and NLRP3 (r = 0.5727, p < 0.05).

    Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

    Techniques: Expressing

    Effect of HHQG on protein expression of RASD1, NLRP3, and GSDMD in CCl4-induced liver injury. (A) Protein expression of RASD1, GSDMD, and NLRP3 in liver tissue and the results of the statistical analysis of the strip gray value. (B) Immunohistochemical staining of RASD1 in liver tissue (200 ×). (C) Immunohistochemical staining of NLRP3 in liver tissue (200 ×). (D) Immunohistochemical staining of GSDMD in liver tissue (200 ×).(Each band in the figure represents a different blot, all samples are from the same experiment, and the blots were processed in parallel.)

    Journal: Scientific Reports

    Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

    doi: 10.1038/s41598-025-24410-z

    Figure Lengend Snippet: Effect of HHQG on protein expression of RASD1, NLRP3, and GSDMD in CCl4-induced liver injury. (A) Protein expression of RASD1, GSDMD, and NLRP3 in liver tissue and the results of the statistical analysis of the strip gray value. (B) Immunohistochemical staining of RASD1 in liver tissue (200 ×). (C) Immunohistochemical staining of NLRP3 in liver tissue (200 ×). (D) Immunohistochemical staining of GSDMD in liver tissue (200 ×).(Each band in the figure represents a different blot, all samples are from the same experiment, and the blots were processed in parallel.)

    Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

    Techniques: Expressing, Stripping Membranes, Immunohistochemical staining, Staining

    Effect of nCGA in decreasing both the pyroptosis and inflammation levels in glucolipotoxicity-induced MES-13 cells. (A) MES-13 cells were subjected to Western blotting to analyze NLRP3, GSDMD, IL-1β, and Caspase-1 expression. (B) Quantification of the protein expression. (C) Annexin-V/PI flow cytometry analysis was performed to detect phosphatidylserine externalization, a marker of both apoptosis and pyroptosis-associated membrane disruption. (D–F) Immunofluorescence staining for the expression and cellular localization of pyroptosis markers. Data are shown as mean ± SD. Results were statistically analyzed using ANOVA, followed by the Bonferroni post-hoc test. * p < 0.05 was considered statistically significant compared to the HG + OA group.

    Journal: Journal of Agricultural and Food Chemistry

    Article Title: Protective Effect of Neochlorogenic Acid on Diabetic Nephropathy via Inflammation and Pyroptosis Suppression

    doi: 10.1021/acs.jafc.5c09087

    Figure Lengend Snippet: Effect of nCGA in decreasing both the pyroptosis and inflammation levels in glucolipotoxicity-induced MES-13 cells. (A) MES-13 cells were subjected to Western blotting to analyze NLRP3, GSDMD, IL-1β, and Caspase-1 expression. (B) Quantification of the protein expression. (C) Annexin-V/PI flow cytometry analysis was performed to detect phosphatidylserine externalization, a marker of both apoptosis and pyroptosis-associated membrane disruption. (D–F) Immunofluorescence staining for the expression and cellular localization of pyroptosis markers. Data are shown as mean ± SD. Results were statistically analyzed using ANOVA, followed by the Bonferroni post-hoc test. * p < 0.05 was considered statistically significant compared to the HG + OA group.

    Article Snippet: Primary antibodies against β-actin (Invitrogen, MA5-11866), NFκb p65 (Santa Cruz, sc-109), p-NFκb (Abclonal, AP0123), IKB (Santa Cruz, sc-847), p-IKB (Cell Signaling, no. 9246), Nrf2 (Santa Cruz, sc-722), TNF-α (Elabscience, E-AB-40015), MCP-1 (Santa Cruz, sc-28879), ICAM-1 (Santa Cruz, sc-107), COX-2 (Santa Cruz, sc-19999), iNOS (Santa Cruz, sc-7271), NLRP3 (Bioss, bs-10021R), GSDMD (Invitrogen, PA5-104324), IL-1β (abcam, ab283818), and Caspase-1 p20 (Bioss, bs-10442R).

    Techniques: Western Blot, Expressing, Flow Cytometry, Marker, Membrane, Disruption, Immunofluorescence, Staining

    Effect of miR-30a overexpression and miR-30a inhibition on pyroptosis in glucolipotoxicity-induced MES-13 cells. (A) Western blot and quantitative analysis of NLRP3, GSDMD-N, IL-1β, and caspase-1 after treatment with HG, OA, nCGA, and miR-30a mimic. (B) Western blot and quantitative analysis of the same proteins after treatment with the HG, OA, nCGA, and miR-30a inhibitor. Data are shown as mean ± SD * p < 0.05 compared to the HG group and # p < 0.05 compared to the HG + OA group.

    Journal: Journal of Agricultural and Food Chemistry

    Article Title: Protective Effect of Neochlorogenic Acid on Diabetic Nephropathy via Inflammation and Pyroptosis Suppression

    doi: 10.1021/acs.jafc.5c09087

    Figure Lengend Snippet: Effect of miR-30a overexpression and miR-30a inhibition on pyroptosis in glucolipotoxicity-induced MES-13 cells. (A) Western blot and quantitative analysis of NLRP3, GSDMD-N, IL-1β, and caspase-1 after treatment with HG, OA, nCGA, and miR-30a mimic. (B) Western blot and quantitative analysis of the same proteins after treatment with the HG, OA, nCGA, and miR-30a inhibitor. Data are shown as mean ± SD * p < 0.05 compared to the HG group and # p < 0.05 compared to the HG + OA group.

    Article Snippet: Primary antibodies against β-actin (Invitrogen, MA5-11866), NFκb p65 (Santa Cruz, sc-109), p-NFκb (Abclonal, AP0123), IKB (Santa Cruz, sc-847), p-IKB (Cell Signaling, no. 9246), Nrf2 (Santa Cruz, sc-722), TNF-α (Elabscience, E-AB-40015), MCP-1 (Santa Cruz, sc-28879), ICAM-1 (Santa Cruz, sc-107), COX-2 (Santa Cruz, sc-19999), iNOS (Santa Cruz, sc-7271), NLRP3 (Bioss, bs-10021R), GSDMD (Invitrogen, PA5-104324), IL-1β (abcam, ab283818), and Caspase-1 p20 (Bioss, bs-10442R).

    Techniques: Over Expression, Inhibition, Western Blot