pf3644022  (Tocris)


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    Tocris pf3644022

    Pf3644022, supplied by Tocris, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pf3644022/product/Tocris
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pf3644022 - by Bioz Stars, 2024-07
    94/100 stars

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    1) Product Images from "Necrosulfonamide causes oxidation of PCM1 and impairs ciliogenesis and autophagy"

    Article Title: Necrosulfonamide causes oxidation of PCM1 and impairs ciliogenesis and autophagy

    Journal: iScience

    doi: 10.1016/j.isci.2024.109580


    Figure Legend Snippet:

    Techniques Used: Recombinant, Transfection, Electron Microscopy, Protease Inhibitor, Western Blot, Purification, SYBR Green Assay, Viability Assay, Sequencing, Software, Imaging

    pf 3644022  (Selleck Chemicals)


    Bioz Manufacturer Symbol Selleck Chemicals manufactures this product  
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    Selleck Chemicals pf 3644022
    Pf 3644022, supplied by Selleck Chemicals, 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/result/pf 3644022/product/Selleck Chemicals
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pf 3644022 - by Bioz Stars, 2024-07
    86/100 stars

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    pf 3644022  (Bio-Techne corporation)


    Bioz Verified Symbol Bio-Techne corporation is a verified supplier
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    Bio-Techne corporation pf 3644022
    Pf 3644022, supplied by Bio-Techne corporation, 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/result/pf 3644022/product/Bio-Techne corporation
    Average 86 stars, based on 1 article reviews
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    86/100 stars

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    SMAC Corp pf 3644022
    Pf 3644022, supplied by SMAC Corp, 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/result/pf 3644022/product/SMAC Corp
    Average 86 stars, based on 1 article reviews
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    pf 3644022 - by Bioz Stars, 2024-07
    86/100 stars

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    pf3644022  (Tocris)


    Bioz Verified Symbol Tocris is a verified supplier
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    Tocris pf3644022

    Pf3644022, supplied by Tocris, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pf3644022/product/Tocris
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pf3644022 - by Bioz Stars, 2024-07
    94/100 stars

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    1) Product Images from "Necrosulfonamide causes oxidation of PCM1 and impairs ciliogenesis and autophagy"

    Article Title: Necrosulfonamide causes oxidation of PCM1 and impairs ciliogenesis and autophagy

    Journal: iScience

    doi: 10.1016/j.isci.2024.109580


    Figure Legend Snippet:

    Techniques Used: Recombinant, Transfection, Electron Microscopy, Protease Inhibitor, Western Blot, Purification, SYBR Green Assay, Viability Assay, Sequencing, Software, Imaging

    mk2 inhibitor pf 3644022  (Selleck Chemicals)


    Bioz Manufacturer Symbol Selleck Chemicals manufactures this product  
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    Selleck Chemicals mk2 inhibitor pf 3644022
    NLRP6 deficiency increases RIP1 phosphorylation at S321 via the TAK1/p38 MAPK <t>/MK2</t> cascade (A) Western blot analysis and relative intensity of HEK293T cells transfected with plasmids encoding His-NLRP6 together with HA-RIP3 indicated concentration of Flag-TAK1. (B) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with the protein synthesis inhibitor CHX for indicated time points. (C) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with S. typhimurium strain SL1344 at an MOI of 10 for 4 h. (D) Immunoblot analysis of whole-cell lysates of 293T cells transfected with plasmids encoding His-NLRP6 and Flag-TAK1, followed by immunoprecipitation with anti-Flag beads. (E) Western blot analysis of protein extracts of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h, followed by immunoprecipitation with anti-TAK1 antibody. (F) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the p38 MAPK inhibitor SB239063 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (G) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (H) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the MK-2 inhibitor MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (I) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (J) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with vehicle or MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. All data are representative of at least three independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.
    Mk2 Inhibitor Pf 3644022, supplied by Selleck Chemicals, 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/result/mk2 inhibitor pf 3644022/product/Selleck Chemicals
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mk2 inhibitor pf 3644022 - by Bioz Stars, 2024-07
    86/100 stars

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    1) Product Images from "NLRP6 induces RIP1 kinase-dependent necroptosis via TAK1-mediated p38 MAPK /MK2 phosphorylation in S . typhimurium infection"

    Article Title: NLRP6 induces RIP1 kinase-dependent necroptosis via TAK1-mediated p38 MAPK /MK2 phosphorylation in S . typhimurium infection

    Journal: iScience

    doi: 10.1016/j.isci.2024.109339

    NLRP6 deficiency increases RIP1 phosphorylation at S321 via the TAK1/p38 MAPK /MK2 cascade (A) Western blot analysis and relative intensity of HEK293T cells transfected with plasmids encoding His-NLRP6 together with HA-RIP3 indicated concentration of Flag-TAK1. (B) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with the protein synthesis inhibitor CHX for indicated time points. (C) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with S. typhimurium strain SL1344 at an MOI of 10 for 4 h. (D) Immunoblot analysis of whole-cell lysates of 293T cells transfected with plasmids encoding His-NLRP6 and Flag-TAK1, followed by immunoprecipitation with anti-Flag beads. (E) Western blot analysis of protein extracts of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h, followed by immunoprecipitation with anti-TAK1 antibody. (F) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the p38 MAPK inhibitor SB239063 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (G) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (H) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the MK-2 inhibitor MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (I) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (J) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with vehicle or MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. All data are representative of at least three independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.
    Figure Legend Snippet: NLRP6 deficiency increases RIP1 phosphorylation at S321 via the TAK1/p38 MAPK /MK2 cascade (A) Western blot analysis and relative intensity of HEK293T cells transfected with plasmids encoding His-NLRP6 together with HA-RIP3 indicated concentration of Flag-TAK1. (B) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with the protein synthesis inhibitor CHX for indicated time points. (C) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with S. typhimurium strain SL1344 at an MOI of 10 for 4 h. (D) Immunoblot analysis of whole-cell lysates of 293T cells transfected with plasmids encoding His-NLRP6 and Flag-TAK1, followed by immunoprecipitation with anti-Flag beads. (E) Western blot analysis of protein extracts of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h, followed by immunoprecipitation with anti-TAK1 antibody. (F) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the p38 MAPK inhibitor SB239063 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (G) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (H) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the MK-2 inhibitor MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (I) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (J) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with vehicle or MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. All data are representative of at least three independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.

    Techniques Used: Western Blot, Transfection, Concentration Assay, Infection, Immunoprecipitation

    NLRP6 deficiency decreases S . typhimurium -induced enterocyte necroptosis and intestinal barrier dysfunction (A) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle, NSA or GSK’872 for 1 h, followed by infection with S . typhimurium strain SL1344 at an MOI of ∼100 for 4 h. (B) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (C) Cell death of WT and NLRP6−/− Caco-2 cells treated with vehicle, zVAD or zVAD + Nec-1 for 1 h, followed by infection with S. typhimurium strain SL1344 at an MOI of ∼100 for 4 h. (D and E) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (F) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle or (5Z)-7-Oxozeaenol for 1 h, followed by infection with SL1344 at an MOI of ∼100 for 4 h. (G) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle or SB239063 for 30 min, followed by infection with SL1344 at an MOI of ∼100 for 4 h. (H) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (I) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle or MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of ∼100 for 4 h. (J) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (K) TEER levels of WT and NLRP6 −/− Caco-2 cell monolayers infected with SL1344 at an MOI of ∼100. (L) FD4 permeability of WT and NLRP6 −/− Caco-2 cell monolayers infected with SL1344 at an MOI of ∼100 for 3 h. (M) Bacterial translocation of WT and NLRP6 −/− Caco-2 cell monolayers infected with SL1344 at an MOI of ∼100 for 3 h. Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. All data are representative of at least three independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001; ns, not significant.
    Figure Legend Snippet: NLRP6 deficiency decreases S . typhimurium -induced enterocyte necroptosis and intestinal barrier dysfunction (A) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle, NSA or GSK’872 for 1 h, followed by infection with S . typhimurium strain SL1344 at an MOI of ∼100 for 4 h. (B) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (C) Cell death of WT and NLRP6−/− Caco-2 cells treated with vehicle, zVAD or zVAD + Nec-1 for 1 h, followed by infection with S. typhimurium strain SL1344 at an MOI of ∼100 for 4 h. (D and E) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (F) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle or (5Z)-7-Oxozeaenol for 1 h, followed by infection with SL1344 at an MOI of ∼100 for 4 h. (G) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle or SB239063 for 30 min, followed by infection with SL1344 at an MOI of ∼100 for 4 h. (H) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (I) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle or MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of ∼100 for 4 h. (J) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (K) TEER levels of WT and NLRP6 −/− Caco-2 cell monolayers infected with SL1344 at an MOI of ∼100. (L) FD4 permeability of WT and NLRP6 −/− Caco-2 cell monolayers infected with SL1344 at an MOI of ∼100 for 3 h. (M) Bacterial translocation of WT and NLRP6 −/− Caco-2 cell monolayers infected with SL1344 at an MOI of ∼100 for 3 h. Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. All data are representative of at least three independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001; ns, not significant.

    Techniques Used: Infection, Western Blot, Permeability, Translocation Assay

    NLRP6 promotes the systemic dissemination of S . typhimurium by regulating MK2/RIP1-associated necroptosis in vivo (A–F) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of S . typhimurium strain SL1344. Mice were administrated intraperitoneally with the RIP1 inhibitor Nec-1s at 1 h prior to infection and 24 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (A, B). Bacterial burden in MLN (C), liver (D) and spleen (E). Intestinal permeability (F). (G and H) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and analyzed at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice. (I–M) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and administrated orally with the MK2 inhibitor PF-3644022 at 4 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (I, J). Bacterial burden in MLN (K), liver (L) and spleen (M). Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.
    Figure Legend Snippet: NLRP6 promotes the systemic dissemination of S . typhimurium by regulating MK2/RIP1-associated necroptosis in vivo (A–F) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of S . typhimurium strain SL1344. Mice were administrated intraperitoneally with the RIP1 inhibitor Nec-1s at 1 h prior to infection and 24 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (A, B). Bacterial burden in MLN (C), liver (D) and spleen (E). Intestinal permeability (F). (G and H) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and analyzed at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice. (I–M) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and administrated orally with the MK2 inhibitor PF-3644022 at 4 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (I, J). Bacterial burden in MLN (K), liver (L) and spleen (M). Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.

    Techniques Used: In Vivo, Infection, Western Blot, Isolation, Permeability


    Figure Legend Snippet:

    Techniques Used: Virus, Recombinant, Modification, Lysis, Lactate Dehydrogenase Assay, Cell Viability Assay, Bradford Protein Assay, Luciferase, Reporter Assay, Western Blot, Sequencing, Plasmid Preparation, Expressing, Software

    pf 3644022  (Selleck Chemicals)


    Bioz Manufacturer Symbol Selleck Chemicals manufactures this product  
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    Selleck Chemicals pf 3644022
    NLRP6 promotes the systemic dissemination of S . typhimurium by regulating MK2/RIP1-associated necroptosis in vivo (A–F) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of S . typhimurium strain SL1344. Mice were administrated intraperitoneally with the RIP1 inhibitor Nec-1s at 1 h prior to infection and 24 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (A, B). Bacterial burden in MLN (C), liver (D) and spleen (E). Intestinal permeability (F). (G and H) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and analyzed at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice. (I–M) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and administrated orally with the MK2 inhibitor <t>PF-3644022</t> at 4 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (I, J). Bacterial burden in MLN (K), liver (L) and spleen (M). Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.
    Pf 3644022, supplied by Selleck Chemicals, 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/result/pf 3644022/product/Selleck Chemicals
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pf 3644022 - by Bioz Stars, 2024-07
    86/100 stars

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    1) Product Images from "NLRP6 induces RIP1 kinase-dependent necroptosis via TAK1-mediated p38 MAPK /MK2 phosphorylation in S . typhimurium infection"

    Article Title: NLRP6 induces RIP1 kinase-dependent necroptosis via TAK1-mediated p38 MAPK /MK2 phosphorylation in S . typhimurium infection

    Journal: iScience

    doi: 10.1016/j.isci.2024.109339

    NLRP6 promotes the systemic dissemination of S . typhimurium by regulating MK2/RIP1-associated necroptosis in vivo (A–F) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of S . typhimurium strain SL1344. Mice were administrated intraperitoneally with the RIP1 inhibitor Nec-1s at 1 h prior to infection and 24 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (A, B). Bacterial burden in MLN (C), liver (D) and spleen (E). Intestinal permeability (F). (G and H) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and analyzed at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice. (I–M) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and administrated orally with the MK2 inhibitor PF-3644022 at 4 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (I, J). Bacterial burden in MLN (K), liver (L) and spleen (M). Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.
    Figure Legend Snippet: NLRP6 promotes the systemic dissemination of S . typhimurium by regulating MK2/RIP1-associated necroptosis in vivo (A–F) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of S . typhimurium strain SL1344. Mice were administrated intraperitoneally with the RIP1 inhibitor Nec-1s at 1 h prior to infection and 24 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (A, B). Bacterial burden in MLN (C), liver (D) and spleen (E). Intestinal permeability (F). (G and H) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and analyzed at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice. (I–M) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and administrated orally with the MK2 inhibitor PF-3644022 at 4 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (I, J). Bacterial burden in MLN (K), liver (L) and spleen (M). Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.

    Techniques Used: In Vivo, Infection, Western Blot, Isolation, Permeability


    Structured Review

    Merck & Co mk2 inhibitor mk2i pf 3644022
    Mk2 Inhibitor Mk2i Pf 3644022, supplied by Merck & Co, 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/result/mk2 inhibitor mk2i pf 3644022/product/Merck & Co
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mk2 inhibitor mk2i pf 3644022 - by Bioz Stars, 2024-07
    86/100 stars

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    Structured Review

    Merck & Co mk2 inhibitor mk2i pf 3644022
    Inhibition of TBK1 results in stimulation of RIPK1-dependent enterocyte shedding. ( A – F ) TBK1i stimulated enterocyte shedding in RV culture, but TAK1i and <t>MK2i</t> did not. RV were treated with 2 µM TBK1i (MRT67307) ( A , B ), 100 or 300 nM TAK1i (5Z-7-Oxozeaenol) ( C , D ), or 1 or 10 µM MK2i <t>(PF-3644022)</t> ( E , F ) for 6 h. In the case of TBK1i, RV were also pre-treated for 30 min with Nec-1s, and then 2 µM TBK1i was added for 6 h. Shed cells were counted; data are the mean ± SEM from three independent experiments ( A , C , E ). In ( B , D , F ), all plots of the three experiments are shown ( B : vehicle, n = 159, TBK1i, n = 163, TBK1i + Nec-1s, n = 151, D : vehicle, n = 142 , TAK1i [100 nM], n = 127, TAK1i [300 nM], n = 130, F : vehicle, n = 181, MK2i [1 µM], n = 151, MK2i [10 µM], n = 167); the horizontal line represents the mean. ( G – I ) The amount of TBK1 and the S321-phosphorylation level of RIPK1 were lower in WT mice housed with αDri than in sex-matched littermate WT mice housed with Eco Chips. ( G ), Western blot analysis of the quantity of TBK1, S321-phosphorylated-RIPK1 (p-RIPK1 [S321]), and total RIPK1 (RIPK1) in the enterocytes of WT mice housed with αDri and sex-matched littermate WT mice housed with Eco Chips. E-cadherin was a loading control. ( H ), Densitometry of TBK1 was normalized to E-cadherin; then, the ratio (relative to a value of Eco Chips as 1) was calculated in each Eco Chips/ αDri pair. The mean ± SEM from three pairs is shown. ( I ), Densitometry was performed to obtain the ratio of p-RIPK1 (S321) to total RIPK1 (normalized to Eco Chips as 1) in each Eco Chips/αDri pair. The mean ± SEM from three pairs is shown. For ( A – F ), P values were calculated using one-way Anova with Dunnett’s multiple comparison tests. For ( H , I ), P values were calculated using one-sample t -tests. * P < 0.05, ** P < 0.01, n.s., not significant. Uncropped Western blot data used in G , H , and I are shown in supplementary Fig. .
    Mk2 Inhibitor Mk2i Pf 3644022, supplied by Merck & Co, 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/result/mk2 inhibitor mk2i pf 3644022/product/Merck & Co
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mk2 inhibitor mk2i pf 3644022 - by Bioz Stars, 2024-07
    86/100 stars

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    1) Product Images from "Housing conditions affect enterocyte death mode and turnover rate in mouse small intestine"

    Article Title: Housing conditions affect enterocyte death mode and turnover rate in mouse small intestine

    Journal: Scientific Reports

    doi: 10.1038/s41598-023-47660-1

    Inhibition of TBK1 results in stimulation of RIPK1-dependent enterocyte shedding. ( A – F ) TBK1i stimulated enterocyte shedding in RV culture, but TAK1i and MK2i did not. RV were treated with 2 µM TBK1i (MRT67307) ( A , B ), 100 or 300 nM TAK1i (5Z-7-Oxozeaenol) ( C , D ), or 1 or 10 µM MK2i (PF-3644022) ( E , F ) for 6 h. In the case of TBK1i, RV were also pre-treated for 30 min with Nec-1s, and then 2 µM TBK1i was added for 6 h. Shed cells were counted; data are the mean ± SEM from three independent experiments ( A , C , E ). In ( B , D , F ), all plots of the three experiments are shown ( B : vehicle, n = 159, TBK1i, n = 163, TBK1i + Nec-1s, n = 151, D : vehicle, n = 142 , TAK1i [100 nM], n = 127, TAK1i [300 nM], n = 130, F : vehicle, n = 181, MK2i [1 µM], n = 151, MK2i [10 µM], n = 167); the horizontal line represents the mean. ( G – I ) The amount of TBK1 and the S321-phosphorylation level of RIPK1 were lower in WT mice housed with αDri than in sex-matched littermate WT mice housed with Eco Chips. ( G ), Western blot analysis of the quantity of TBK1, S321-phosphorylated-RIPK1 (p-RIPK1 [S321]), and total RIPK1 (RIPK1) in the enterocytes of WT mice housed with αDri and sex-matched littermate WT mice housed with Eco Chips. E-cadherin was a loading control. ( H ), Densitometry of TBK1 was normalized to E-cadherin; then, the ratio (relative to a value of Eco Chips as 1) was calculated in each Eco Chips/ αDri pair. The mean ± SEM from three pairs is shown. ( I ), Densitometry was performed to obtain the ratio of p-RIPK1 (S321) to total RIPK1 (normalized to Eco Chips as 1) in each Eco Chips/αDri pair. The mean ± SEM from three pairs is shown. For ( A – F ), P values were calculated using one-way Anova with Dunnett’s multiple comparison tests. For ( H , I ), P values were calculated using one-sample t -tests. * P < 0.05, ** P < 0.01, n.s., not significant. Uncropped Western blot data used in G , H , and I are shown in supplementary Fig. .
    Figure Legend Snippet: Inhibition of TBK1 results in stimulation of RIPK1-dependent enterocyte shedding. ( A – F ) TBK1i stimulated enterocyte shedding in RV culture, but TAK1i and MK2i did not. RV were treated with 2 µM TBK1i (MRT67307) ( A , B ), 100 or 300 nM TAK1i (5Z-7-Oxozeaenol) ( C , D ), or 1 or 10 µM MK2i (PF-3644022) ( E , F ) for 6 h. In the case of TBK1i, RV were also pre-treated for 30 min with Nec-1s, and then 2 µM TBK1i was added for 6 h. Shed cells were counted; data are the mean ± SEM from three independent experiments ( A , C , E ). In ( B , D , F ), all plots of the three experiments are shown ( B : vehicle, n = 159, TBK1i, n = 163, TBK1i + Nec-1s, n = 151, D : vehicle, n = 142 , TAK1i [100 nM], n = 127, TAK1i [300 nM], n = 130, F : vehicle, n = 181, MK2i [1 µM], n = 151, MK2i [10 µM], n = 167); the horizontal line represents the mean. ( G – I ) The amount of TBK1 and the S321-phosphorylation level of RIPK1 were lower in WT mice housed with αDri than in sex-matched littermate WT mice housed with Eco Chips. ( G ), Western blot analysis of the quantity of TBK1, S321-phosphorylated-RIPK1 (p-RIPK1 [S321]), and total RIPK1 (RIPK1) in the enterocytes of WT mice housed with αDri and sex-matched littermate WT mice housed with Eco Chips. E-cadherin was a loading control. ( H ), Densitometry of TBK1 was normalized to E-cadherin; then, the ratio (relative to a value of Eco Chips as 1) was calculated in each Eco Chips/ αDri pair. The mean ± SEM from three pairs is shown. ( I ), Densitometry was performed to obtain the ratio of p-RIPK1 (S321) to total RIPK1 (normalized to Eco Chips as 1) in each Eco Chips/αDri pair. The mean ± SEM from three pairs is shown. For ( A – F ), P values were calculated using one-way Anova with Dunnett’s multiple comparison tests. For ( H , I ), P values were calculated using one-sample t -tests. * P < 0.05, ** P < 0.01, n.s., not significant. Uncropped Western blot data used in G , H , and I are shown in supplementary Fig. .

    Techniques Used: Inhibition, Western Blot, Comparison

    pf  (Tocris)


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    Tocris pf3644022

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    NLRP6 deficiency increases RIP1 phosphorylation at S321 via the TAK1/p38 MAPK <t>/MK2</t> cascade (A) Western blot analysis and relative intensity of HEK293T cells transfected with plasmids encoding His-NLRP6 together with HA-RIP3 indicated concentration of Flag-TAK1. (B) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with the protein synthesis inhibitor CHX for indicated time points. (C) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with S. typhimurium strain SL1344 at an MOI of 10 for 4 h. (D) Immunoblot analysis of whole-cell lysates of 293T cells transfected with plasmids encoding His-NLRP6 and Flag-TAK1, followed by immunoprecipitation with anti-Flag beads. (E) Western blot analysis of protein extracts of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h, followed by immunoprecipitation with anti-TAK1 antibody. (F) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the p38 MAPK inhibitor SB239063 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (G) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (H) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the MK-2 inhibitor MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (I) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (J) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with vehicle or MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. All data are representative of at least three independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.
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    NLRP6 deficiency increases RIP1 phosphorylation at S321 via the TAK1/p38 MAPK <t>/MK2</t> cascade (A) Western blot analysis and relative intensity of HEK293T cells transfected with plasmids encoding His-NLRP6 together with HA-RIP3 indicated concentration of Flag-TAK1. (B) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with the protein synthesis inhibitor CHX for indicated time points. (C) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with S. typhimurium strain SL1344 at an MOI of 10 for 4 h. (D) Immunoblot analysis of whole-cell lysates of 293T cells transfected with plasmids encoding His-NLRP6 and Flag-TAK1, followed by immunoprecipitation with anti-Flag beads. (E) Western blot analysis of protein extracts of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h, followed by immunoprecipitation with anti-TAK1 antibody. (F) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the p38 MAPK inhibitor SB239063 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (G) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (H) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the MK-2 inhibitor MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (I) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (J) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with vehicle or MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. All data are representative of at least three independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.
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    NLRP6 deficiency increases RIP1 phosphorylation at S321 via the TAK1/p38 MAPK <t>/MK2</t> cascade (A) Western blot analysis and relative intensity of HEK293T cells transfected with plasmids encoding His-NLRP6 together with HA-RIP3 indicated concentration of Flag-TAK1. (B) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with the protein synthesis inhibitor CHX for indicated time points. (C) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with S. typhimurium strain SL1344 at an MOI of 10 for 4 h. (D) Immunoblot analysis of whole-cell lysates of 293T cells transfected with plasmids encoding His-NLRP6 and Flag-TAK1, followed by immunoprecipitation with anti-Flag beads. (E) Western blot analysis of protein extracts of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h, followed by immunoprecipitation with anti-TAK1 antibody. (F) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the p38 MAPK inhibitor SB239063 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (G) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (H) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the MK-2 inhibitor MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (I) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (J) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with vehicle or MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. All data are representative of at least three independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.
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    NLRP6 deficiency increases RIP1 phosphorylation at S321 via the TAK1/p38 MAPK <t>/MK2</t> cascade (A) Western blot analysis and relative intensity of HEK293T cells transfected with plasmids encoding His-NLRP6 together with HA-RIP3 indicated concentration of Flag-TAK1. (B) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with the protein synthesis inhibitor CHX for indicated time points. (C) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with S. typhimurium strain SL1344 at an MOI of 10 for 4 h. (D) Immunoblot analysis of whole-cell lysates of 293T cells transfected with plasmids encoding His-NLRP6 and Flag-TAK1, followed by immunoprecipitation with anti-Flag beads. (E) Western blot analysis of protein extracts of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h, followed by immunoprecipitation with anti-TAK1 antibody. (F) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the p38 MAPK inhibitor SB239063 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (G) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (H) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the MK-2 inhibitor MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (I) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (J) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with vehicle or MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. All data are representative of at least three independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.
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    Journal: iScience

    Article Title: Necrosulfonamide causes oxidation of PCM1 and impairs ciliogenesis and autophagy

    doi: 10.1016/j.isci.2024.109580

    Figure Lengend Snippet:

    Article Snippet: PF3644022 , Tocris , Cat#4279.

    Techniques: Recombinant, Transfection, Electron Microscopy, Protease Inhibitor, Western Blot, Purification, SYBR Green Assay, Viability Assay, Sequencing, Software, Imaging

    NLRP6 deficiency increases RIP1 phosphorylation at S321 via the TAK1/p38 MAPK /MK2 cascade (A) Western blot analysis and relative intensity of HEK293T cells transfected with plasmids encoding His-NLRP6 together with HA-RIP3 indicated concentration of Flag-TAK1. (B) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with the protein synthesis inhibitor CHX for indicated time points. (C) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with S. typhimurium strain SL1344 at an MOI of 10 for 4 h. (D) Immunoblot analysis of whole-cell lysates of 293T cells transfected with plasmids encoding His-NLRP6 and Flag-TAK1, followed by immunoprecipitation with anti-Flag beads. (E) Western blot analysis of protein extracts of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h, followed by immunoprecipitation with anti-TAK1 antibody. (F) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the p38 MAPK inhibitor SB239063 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (G) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (H) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the MK-2 inhibitor MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (I) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (J) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with vehicle or MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. All data are representative of at least three independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.

    Journal: iScience

    Article Title: NLRP6 induces RIP1 kinase-dependent necroptosis via TAK1-mediated p38 MAPK /MK2 phosphorylation in S . typhimurium infection

    doi: 10.1016/j.isci.2024.109339

    Figure Lengend Snippet: NLRP6 deficiency increases RIP1 phosphorylation at S321 via the TAK1/p38 MAPK /MK2 cascade (A) Western blot analysis and relative intensity of HEK293T cells transfected with plasmids encoding His-NLRP6 together with HA-RIP3 indicated concentration of Flag-TAK1. (B) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with the protein synthesis inhibitor CHX for indicated time points. (C) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with S. typhimurium strain SL1344 at an MOI of 10 for 4 h. (D) Immunoblot analysis of whole-cell lysates of 293T cells transfected with plasmids encoding His-NLRP6 and Flag-TAK1, followed by immunoprecipitation with anti-Flag beads. (E) Western blot analysis of protein extracts of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h, followed by immunoprecipitation with anti-TAK1 antibody. (F) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the p38 MAPK inhibitor SB239063 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (G) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (H) Cell death of WT and Nlrp6 −/− MEFs treated with either vehicle or the MK-2 inhibitor MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. (I) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs infected with SL1344 at an MOI of 10 for 4 h. (J) Western blot analysis and relative intensity of WT and Nlrp6 −/− MEFs treated with vehicle or MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of 10 for 4 h. Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. All data are representative of at least three independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.

    Article Snippet: When appropriate, mice were treated with the RIP1 inhibitor Nec-1s (S8641, Selleck Chemicals, Houston, TX, USA) or the MK2 inhibitor PF-3644022 (S8224, Selleck Chemicals).

    Techniques: Western Blot, Transfection, Concentration Assay, Infection, Immunoprecipitation

    NLRP6 deficiency decreases S . typhimurium -induced enterocyte necroptosis and intestinal barrier dysfunction (A) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle, NSA or GSK’872 for 1 h, followed by infection with S . typhimurium strain SL1344 at an MOI of ∼100 for 4 h. (B) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (C) Cell death of WT and NLRP6−/− Caco-2 cells treated with vehicle, zVAD or zVAD + Nec-1 for 1 h, followed by infection with S. typhimurium strain SL1344 at an MOI of ∼100 for 4 h. (D and E) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (F) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle or (5Z)-7-Oxozeaenol for 1 h, followed by infection with SL1344 at an MOI of ∼100 for 4 h. (G) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle or SB239063 for 30 min, followed by infection with SL1344 at an MOI of ∼100 for 4 h. (H) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (I) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle or MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of ∼100 for 4 h. (J) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (K) TEER levels of WT and NLRP6 −/− Caco-2 cell monolayers infected with SL1344 at an MOI of ∼100. (L) FD4 permeability of WT and NLRP6 −/− Caco-2 cell monolayers infected with SL1344 at an MOI of ∼100 for 3 h. (M) Bacterial translocation of WT and NLRP6 −/− Caco-2 cell monolayers infected with SL1344 at an MOI of ∼100 for 3 h. Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. All data are representative of at least three independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001; ns, not significant.

    Journal: iScience

    Article Title: NLRP6 induces RIP1 kinase-dependent necroptosis via TAK1-mediated p38 MAPK /MK2 phosphorylation in S . typhimurium infection

    doi: 10.1016/j.isci.2024.109339

    Figure Lengend Snippet: NLRP6 deficiency decreases S . typhimurium -induced enterocyte necroptosis and intestinal barrier dysfunction (A) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle, NSA or GSK’872 for 1 h, followed by infection with S . typhimurium strain SL1344 at an MOI of ∼100 for 4 h. (B) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (C) Cell death of WT and NLRP6−/− Caco-2 cells treated with vehicle, zVAD or zVAD + Nec-1 for 1 h, followed by infection with S. typhimurium strain SL1344 at an MOI of ∼100 for 4 h. (D and E) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (F) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle or (5Z)-7-Oxozeaenol for 1 h, followed by infection with SL1344 at an MOI of ∼100 for 4 h. (G) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle or SB239063 for 30 min, followed by infection with SL1344 at an MOI of ∼100 for 4 h. (H) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (I) Cell death of WT and NLRP6 −/− Caco-2 cells treated with vehicle or MK2-IN-1 for 30 min, followed by infection with SL1344 at an MOI of ∼100 for 4 h. (J) Western blot analysis and relative intensity of WT and NLRP6 −/− Caco-2 cells infected with SL1344 at an MOI of ∼100 for 4 h. (K) TEER levels of WT and NLRP6 −/− Caco-2 cell monolayers infected with SL1344 at an MOI of ∼100. (L) FD4 permeability of WT and NLRP6 −/− Caco-2 cell monolayers infected with SL1344 at an MOI of ∼100 for 3 h. (M) Bacterial translocation of WT and NLRP6 −/− Caco-2 cell monolayers infected with SL1344 at an MOI of ∼100 for 3 h. Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. All data are representative of at least three independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001; ns, not significant.

    Article Snippet: When appropriate, mice were treated with the RIP1 inhibitor Nec-1s (S8641, Selleck Chemicals, Houston, TX, USA) or the MK2 inhibitor PF-3644022 (S8224, Selleck Chemicals).

    Techniques: Infection, Western Blot, Permeability, Translocation Assay

    NLRP6 promotes the systemic dissemination of S . typhimurium by regulating MK2/RIP1-associated necroptosis in vivo (A–F) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of S . typhimurium strain SL1344. Mice were administrated intraperitoneally with the RIP1 inhibitor Nec-1s at 1 h prior to infection and 24 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (A, B). Bacterial burden in MLN (C), liver (D) and spleen (E). Intestinal permeability (F). (G and H) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and analyzed at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice. (I–M) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and administrated orally with the MK2 inhibitor PF-3644022 at 4 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (I, J). Bacterial burden in MLN (K), liver (L) and spleen (M). Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.

    Journal: iScience

    Article Title: NLRP6 induces RIP1 kinase-dependent necroptosis via TAK1-mediated p38 MAPK /MK2 phosphorylation in S . typhimurium infection

    doi: 10.1016/j.isci.2024.109339

    Figure Lengend Snippet: NLRP6 promotes the systemic dissemination of S . typhimurium by regulating MK2/RIP1-associated necroptosis in vivo (A–F) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of S . typhimurium strain SL1344. Mice were administrated intraperitoneally with the RIP1 inhibitor Nec-1s at 1 h prior to infection and 24 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (A, B). Bacterial burden in MLN (C), liver (D) and spleen (E). Intestinal permeability (F). (G and H) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and analyzed at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice. (I–M) Streptomycin-pretreated WT and Nlrp6 −/− mice were orally infected with 1 × 10 8 CFU of SL1344 and administrated orally with the MK2 inhibitor PF-3644022 at 4 h post-infection, followed by sample collection at 2 days post-infection. Western blot analysis and relative intensity of colonic intestinal epithelial cells isolated from mice (I, J). Bacterial burden in MLN (K), liver (L) and spleen (M). Data were compared with independent Student’s t test. Values are expressed as the mean ± SEM, and statistically significant differences are indicated. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.

    Article Snippet: When appropriate, mice were treated with the RIP1 inhibitor Nec-1s (S8641, Selleck Chemicals, Houston, TX, USA) or the MK2 inhibitor PF-3644022 (S8224, Selleck Chemicals).

    Techniques: In Vivo, Infection, Western Blot, Isolation, Permeability

    Journal: iScience

    Article Title: NLRP6 induces RIP1 kinase-dependent necroptosis via TAK1-mediated p38 MAPK /MK2 phosphorylation in S . typhimurium infection

    doi: 10.1016/j.isci.2024.109339

    Figure Lengend Snippet:

    Article Snippet: When appropriate, mice were treated with the RIP1 inhibitor Nec-1s (S8641, Selleck Chemicals, Houston, TX, USA) or the MK2 inhibitor PF-3644022 (S8224, Selleck Chemicals).

    Techniques: Virus, Recombinant, Modification, Lysis, Lactate Dehydrogenase Assay, Cell Viability Assay, Bradford Protein Assay, Luciferase, Reporter Assay, Western Blot, Sequencing, Plasmid Preparation, Expressing, Software