liberase enzymatic digestion Search Results


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  • 99
    Worthington Biochemical dnase
    Dnase, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 99/100, based on 1650 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dnase/product/Worthington Biochemical
    Average 99 stars, based on 1650 article reviews
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    Thermo Fisher hepes
    Hepes, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 52140 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 52140 article reviews
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    hepes - by Bioz Stars, 2020-09
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    99
    Millipore dnase i
    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
    Dnase I, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 34031 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dnase i/product/Millipore
    Average 99 stars, based on 34031 article reviews
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    dnase i - by Bioz Stars, 2020-09
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    Millipore dnase
    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
    Dnase, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 15851 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dnase/product/Millipore
    Average 99 stars, based on 15851 article reviews
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    dnase - by Bioz Stars, 2020-09
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    99
    Millipore liberase
    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
    Liberase, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 723 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/liberase/product/Millipore
    Average 99 stars, based on 723 article reviews
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    liberase - by Bioz Stars, 2020-09
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    99
    Millipore hyaluronidase
    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
    Hyaluronidase, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 12560 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hyaluronidase/product/Millipore
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    99
    Thermo Fisher dmem
    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
    Dmem, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 147955 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    dmem - by Bioz Stars, 2020-09
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    Thermo Fisher fetal bovine serum
    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
    Fetal Bovine Serum, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 225914 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    fetal bovine serum - by Bioz Stars, 2020-09
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    99
    Thermo Fisher trypsin
    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
    Trypsin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 30400 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore bsa
    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
    Bsa, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 42788 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    bsa - by Bioz Stars, 2020-09
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    95
    Millipore liberase dh
    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
    Liberase Dh, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/liberase dh/product/Millipore
    Average 95 stars, based on 94 article reviews
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    liberase dh - by Bioz Stars, 2020-09
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    99
    Thermo Fisher gentamicin sulfate
    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
    Gentamicin Sulfate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 734 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    gentamicin sulfate - by Bioz Stars, 2020-09
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    99
    Millipore glutamine
    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
    Glutamine, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 5582 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glutamine/product/Millipore
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    99
    Thermo Fisher l glutamine
    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
    L Glutamine, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 146399 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson cm2 flask
    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
    Cm2 Flask, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 133 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore elastase
    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
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    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
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    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
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    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
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    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
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    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular <t>DNase</t> I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p
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    Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular DNase I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p

    Journal: Nature Communications

    Article Title: STING-dependent sensing of self-DNA drives silica-induced lung inflammation

    doi: 10.1038/s41467-018-07425-1

    Figure Lengend Snippet: Extracellular self-dsDNA is key to silica-induced inflammatory response in macrophages, not in DCs. a – c Extracellular DNase I treatment (1 µg/mL) was applied 3 h prior and 1 h after silica exposure (250 µg/mL) or transfection with c-di-AMP (6 µg/mL; cDN) for 18 h in bone marrow-derived macrophages ( b ) and dentritic cells ( c ). b Macrophage concentration of extracellular dsDNA and CXCL10 in culture supernatant. Ifnα and Ifnβ transcripts measured by real-time PCR on cell fractions and IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. c Dendritic cell IFN-α and IFN-β protein concentrations determined in culture supernatants by multiplex immunoassay. d – i Mitochondrial DNA replication was inhibited using a low concentration of EtdBr (150 ng/mL) on day 7 of bone marrow cell culture. On day 11, bone marrow-derived DCs ( e – h ) and macrophages ( i ) were unstimulated, stimulated with silica (250 µg/mL), or transfected with c-di-AMP (6 µg/mL; cDN) for 18 h. Fold change of ( e ) mitochondrial DNA (mMitoF1 and mMitoR1) and ( f ) nuclear DNA (mB2MF1 and mB2MR1) in untreated versus EtdBr-treated DC exposed to silica, as compared to untreated unstimulated cells. g Immunoblot showing cGAS, phospho-STING, STING protein expression, and dimerization in DCs after EtdBr treatment, with β-actin as a reference. h CXCL10 level in DC supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. i CXCL10 level in macrophage supernatant quantified by ELISA and IFN-α and IFN-β concentrations determined by multiplex immunoassay. Tmem173 transcripts measured by real-time PCR. * p

    Article Snippet: Flow cytometry The left lung lobe was harvested and lung cells were isolated by enzymatic digestion using liberase (20 μg/mL; Roche) and DNase I (5 mg/mL; Sigma-Aldrich) in a final volume of 5 mL of RPMI at 37°C for 30 min under agitation.

    Techniques: Transfection, Derivative Assay, Concentration Assay, Real-time Polymerase Chain Reaction, Multiplex Assay, Cell Culture, Expressing, Enzyme-linked Immunosorbent Assay

    Self-dsDNA release is central to silica-induced lung inflammation and type I IFN response. a – m Silica microparticles (1 mg/mouse, i.t.) or saline were administered in WT mice and parameters were analyzed on day 7. a Concentration of extracellular dsDNA in the acellular fractions of bronchoalveolar lavage fluid (BALF). b – d Tmem173 , Mb21d1, Ifnα, and Ifnβ transcripts in the lungs and normalized to Gapdh expression. e Lung CXCL10 determined by ELISA and IFN-αβ proteins quantified by multiplex immunoassay. f Correlation between extracellular dsDNA concentrations and type I IFN gene expression. g Annexin V/PI flow cytometry analysis pre-gated on singlet cells. h Correlation between dead cells and extracellular dsDNA. i Increased nuclear DNA (nDNA) and mitochondrial DNA (mtDNA) in the BALF after silica exposure. j Immunoblots of caspase 3 (Casp3), cleaved caspase 3 (c-Casp3), gasdermin D (GSDMD), MLKL, and phosphorylated-MLKL (p-MLKL), normalized to β-actin. Immunoblot quantifications of ( k ) c-Casp3, ( l ) c-GSDMD, and ( m ) p-MLKL. n – u Silica microparticles administered with DNase I (200 µg/mouse, i.p.) as indicated ( n ). o Extracellular dsDNA in BALF acellular fraction. p Lung immunoblots of phospho-STING, STING, phospho-TBK1, TBK1, phospho-IRF3, IRF3, caspase 3 (Casp3), cleaved caspase 3 (c-Casp3), MLKL, and phosphorylated-MLKL (p-MLKL), with β-actin as a reference. q Quantification of STING dimer, relative to β-actin. r STING immunoblots under 5% reducing (left) or 1% 2-mercaptoethanol seminative conditions (2-ME; right). s Lung confocal images of DNA dye Draq5 (cyan) and STING (red). Bars, 20 µm. t Pulmonary IFN-αβ and CXCL10. u Neutrophils, macrophages, and protein extravasation in the BALF. * p

    Journal: Nature Communications

    Article Title: STING-dependent sensing of self-DNA drives silica-induced lung inflammation

    doi: 10.1038/s41467-018-07425-1

    Figure Lengend Snippet: Self-dsDNA release is central to silica-induced lung inflammation and type I IFN response. a – m Silica microparticles (1 mg/mouse, i.t.) or saline were administered in WT mice and parameters were analyzed on day 7. a Concentration of extracellular dsDNA in the acellular fractions of bronchoalveolar lavage fluid (BALF). b – d Tmem173 , Mb21d1, Ifnα, and Ifnβ transcripts in the lungs and normalized to Gapdh expression. e Lung CXCL10 determined by ELISA and IFN-αβ proteins quantified by multiplex immunoassay. f Correlation between extracellular dsDNA concentrations and type I IFN gene expression. g Annexin V/PI flow cytometry analysis pre-gated on singlet cells. h Correlation between dead cells and extracellular dsDNA. i Increased nuclear DNA (nDNA) and mitochondrial DNA (mtDNA) in the BALF after silica exposure. j Immunoblots of caspase 3 (Casp3), cleaved caspase 3 (c-Casp3), gasdermin D (GSDMD), MLKL, and phosphorylated-MLKL (p-MLKL), normalized to β-actin. Immunoblot quantifications of ( k ) c-Casp3, ( l ) c-GSDMD, and ( m ) p-MLKL. n – u Silica microparticles administered with DNase I (200 µg/mouse, i.p.) as indicated ( n ). o Extracellular dsDNA in BALF acellular fraction. p Lung immunoblots of phospho-STING, STING, phospho-TBK1, TBK1, phospho-IRF3, IRF3, caspase 3 (Casp3), cleaved caspase 3 (c-Casp3), MLKL, and phosphorylated-MLKL (p-MLKL), with β-actin as a reference. q Quantification of STING dimer, relative to β-actin. r STING immunoblots under 5% reducing (left) or 1% 2-mercaptoethanol seminative conditions (2-ME; right). s Lung confocal images of DNA dye Draq5 (cyan) and STING (red). Bars, 20 µm. t Pulmonary IFN-αβ and CXCL10. u Neutrophils, macrophages, and protein extravasation in the BALF. * p

    Article Snippet: Flow cytometry The left lung lobe was harvested and lung cells were isolated by enzymatic digestion using liberase (20 μg/mL; Roche) and DNase I (5 mg/mL; Sigma-Aldrich) in a final volume of 5 mL of RPMI at 37°C for 30 min under agitation.

    Techniques: Mouse Assay, Concentration Assay, Expressing, Enzyme-linked Immunosorbent Assay, Multiplex Assay, Flow Cytometry, Cytometry, Western Blot