control-oriented dynamic model of pem fuel cell Search Results


incucyte live cell analysis systems  (Sartorius AG)
99
Sartorius AG incucyte live cell analysis systems
a , b Immunoblot analysis of TNF-RSC isolated from ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cell lines. Cells were stimulated with SF-TNF (500 ng/ml) as indicated, and lysates were subjected to anti-Flag immunoprecipitation. As a control, SF-TNF was added post-lysis to unstimulated samples. A representative experiment ( a ) and quantification of recruitment of indicated proteins to TNF-RSC from three independent experiments normalized to WT cells, mean + SEM ( b ). c , d Immunoblot analysis of lysates of ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cells stimulated with TNF (100 ng/ml) as indicated. A representative experiment ( c ) and quantification of three independent experiments normalized to the maximal response of WT cells, mean ± SEM ( d ). Statistical analysis was based on the area under the curve (AUC) for particular cell lines in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. e , f Immunoblot analysis of lysates from ST2 WT or Tank/Azi2 DKO cells that were unstimulated or stimulated for 24 h with TNF (250 ng/ml) and zVAD (20 µM) as indicated and subjected to RIPK1 immunoprecipitation. A representative experiment ( e ) and quantification of phosphorylated RIPK1 and cleaved Caspase-8 from three independent experiments normalized to WT cells stimulated with TNF alone, mean + SEM ( f ). g ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml), and the induction of cell death was monitored every 2 h using the <t>Incucyte</t> imaging system. The mean ± SEM from six separate experiments using two different Tank/Azi2 DKO ST2 clones is shown. Statistical analysis was based on the AUC for different cell lines and treatments in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. h ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml) alone or in the presence of zVAD (20 µM), Nec-1s (20 µM), or both inhibitors. Induction of cell death after 48 h was measured using the Incucyte imaging system. The mean + SEM from six independent experiments using two different Tank/Azi2 DKO ST2 clones is shown, two-way ANOVA with Tukey’s post-tests. n.s., not significant. #, nonspecific band.
Incucyte Live Cell Analysis Systems, supplied by Sartorius AG, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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uricase colorimetric test  (Thermo Fisher)
90
Thermo Fisher uricase colorimetric test
a , b Immunoblot analysis of TNF-RSC isolated from ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cell lines. Cells were stimulated with SF-TNF (500 ng/ml) as indicated, and lysates were subjected to anti-Flag immunoprecipitation. As a control, SF-TNF was added post-lysis to unstimulated samples. A representative experiment ( a ) and quantification of recruitment of indicated proteins to TNF-RSC from three independent experiments normalized to WT cells, mean + SEM ( b ). c , d Immunoblot analysis of lysates of ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cells stimulated with TNF (100 ng/ml) as indicated. A representative experiment ( c ) and quantification of three independent experiments normalized to the maximal response of WT cells, mean ± SEM ( d ). Statistical analysis was based on the area under the curve (AUC) for particular cell lines in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. e , f Immunoblot analysis of lysates from ST2 WT or Tank/Azi2 DKO cells that were unstimulated or stimulated for 24 h with TNF (250 ng/ml) and zVAD (20 µM) as indicated and subjected to RIPK1 immunoprecipitation. A representative experiment ( e ) and quantification of phosphorylated RIPK1 and cleaved Caspase-8 from three independent experiments normalized to WT cells stimulated with TNF alone, mean + SEM ( f ). g ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml), and the induction of cell death was monitored every 2 h using the <t>Incucyte</t> imaging system. The mean ± SEM from six separate experiments using two different Tank/Azi2 DKO ST2 clones is shown. Statistical analysis was based on the AUC for different cell lines and treatments in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. h ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml) alone or in the presence of zVAD (20 µM), Nec-1s (20 µM), or both inhibitors. Induction of cell death after 48 h was measured using the Incucyte imaging system. The mean + SEM from six independent experiments using two different Tank/Azi2 DKO ST2 clones is shown, two-way ANOVA with Tukey’s post-tests. n.s., not significant. #, nonspecific band.
Uricase Colorimetric Test, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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software package matlab/simulink  (MathWorks Inc)
90
MathWorks Inc software package matlab/simulink
a , b Immunoblot analysis of TNF-RSC isolated from ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cell lines. Cells were stimulated with SF-TNF (500 ng/ml) as indicated, and lysates were subjected to anti-Flag immunoprecipitation. As a control, SF-TNF was added post-lysis to unstimulated samples. A representative experiment ( a ) and quantification of recruitment of indicated proteins to TNF-RSC from three independent experiments normalized to WT cells, mean + SEM ( b ). c , d Immunoblot analysis of lysates of ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cells stimulated with TNF (100 ng/ml) as indicated. A representative experiment ( c ) and quantification of three independent experiments normalized to the maximal response of WT cells, mean ± SEM ( d ). Statistical analysis was based on the area under the curve (AUC) for particular cell lines in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. e , f Immunoblot analysis of lysates from ST2 WT or Tank/Azi2 DKO cells that were unstimulated or stimulated for 24 h with TNF (250 ng/ml) and zVAD (20 µM) as indicated and subjected to RIPK1 immunoprecipitation. A representative experiment ( e ) and quantification of phosphorylated RIPK1 and cleaved Caspase-8 from three independent experiments normalized to WT cells stimulated with TNF alone, mean + SEM ( f ). g ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml), and the induction of cell death was monitored every 2 h using the <t>Incucyte</t> imaging system. The mean ± SEM from six separate experiments using two different Tank/Azi2 DKO ST2 clones is shown. Statistical analysis was based on the AUC for different cell lines and treatments in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. h ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml) alone or in the presence of zVAD (20 µM), Nec-1s (20 µM), or both inhibitors. Induction of cell death after 48 h was measured using the Incucyte imaging system. The mean + SEM from six independent experiments using two different Tank/Azi2 DKO ST2 clones is shown, two-way ANOVA with Tukey’s post-tests. n.s., not significant. #, nonspecific band.
Software Package Matlab/Simulink, supplied by MathWorks Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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sta 409 pc luxx  (NETZSCH)
90
NETZSCH sta 409 pc luxx
a , b Immunoblot analysis of TNF-RSC isolated from ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cell lines. Cells were stimulated with SF-TNF (500 ng/ml) as indicated, and lysates were subjected to anti-Flag immunoprecipitation. As a control, SF-TNF was added post-lysis to unstimulated samples. A representative experiment ( a ) and quantification of recruitment of indicated proteins to TNF-RSC from three independent experiments normalized to WT cells, mean + SEM ( b ). c , d Immunoblot analysis of lysates of ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cells stimulated with TNF (100 ng/ml) as indicated. A representative experiment ( c ) and quantification of three independent experiments normalized to the maximal response of WT cells, mean ± SEM ( d ). Statistical analysis was based on the area under the curve (AUC) for particular cell lines in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. e , f Immunoblot analysis of lysates from ST2 WT or Tank/Azi2 DKO cells that were unstimulated or stimulated for 24 h with TNF (250 ng/ml) and zVAD (20 µM) as indicated and subjected to RIPK1 immunoprecipitation. A representative experiment ( e ) and quantification of phosphorylated RIPK1 and cleaved Caspase-8 from three independent experiments normalized to WT cells stimulated with TNF alone, mean + SEM ( f ). g ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml), and the induction of cell death was monitored every 2 h using the <t>Incucyte</t> imaging system. The mean ± SEM from six separate experiments using two different Tank/Azi2 DKO ST2 clones is shown. Statistical analysis was based on the AUC for different cell lines and treatments in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. h ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml) alone or in the presence of zVAD (20 µM), Nec-1s (20 µM), or both inhibitors. Induction of cell death after 48 h was measured using the Incucyte imaging system. The mean + SEM from six independent experiments using two different Tank/Azi2 DKO ST2 clones is shown, two-way ANOVA with Tukey’s post-tests. n.s., not significant. #, nonspecific band.
Sta 409 Pc Luxx, supplied by NETZSCH, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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171 analysis systems  (Bio-Rad)
93
Bio-Rad 171 analysis systems
a , b Immunoblot analysis of TNF-RSC isolated from ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cell lines. Cells were stimulated with SF-TNF (500 ng/ml) as indicated, and lysates were subjected to anti-Flag immunoprecipitation. As a control, SF-TNF was added post-lysis to unstimulated samples. A representative experiment ( a ) and quantification of recruitment of indicated proteins to TNF-RSC from three independent experiments normalized to WT cells, mean + SEM ( b ). c , d Immunoblot analysis of lysates of ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cells stimulated with TNF (100 ng/ml) as indicated. A representative experiment ( c ) and quantification of three independent experiments normalized to the maximal response of WT cells, mean ± SEM ( d ). Statistical analysis was based on the area under the curve (AUC) for particular cell lines in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. e , f Immunoblot analysis of lysates from ST2 WT or Tank/Azi2 DKO cells that were unstimulated or stimulated for 24 h with TNF (250 ng/ml) and zVAD (20 µM) as indicated and subjected to RIPK1 immunoprecipitation. A representative experiment ( e ) and quantification of phosphorylated RIPK1 and cleaved Caspase-8 from three independent experiments normalized to WT cells stimulated with TNF alone, mean + SEM ( f ). g ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml), and the induction of cell death was monitored every 2 h using the <t>Incucyte</t> imaging system. The mean ± SEM from six separate experiments using two different Tank/Azi2 DKO ST2 clones is shown. Statistical analysis was based on the AUC for different cell lines and treatments in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. h ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml) alone or in the presence of zVAD (20 µM), Nec-1s (20 µM), or both inhibitors. Induction of cell death after 48 h was measured using the Incucyte imaging system. The mean + SEM from six independent experiments using two different Tank/Azi2 DKO ST2 clones is shown, two-way ANOVA with Tukey’s post-tests. n.s., not significant. #, nonspecific band.
171 Analysis Systems, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rapid analyte measurement platform (rampt) wnv test  (Response Biomedical)
90
Response Biomedical rapid analyte measurement platform (rampt) wnv test
a , b Immunoblot analysis of TNF-RSC isolated from ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cell lines. Cells were stimulated with SF-TNF (500 ng/ml) as indicated, and lysates were subjected to anti-Flag immunoprecipitation. As a control, SF-TNF was added post-lysis to unstimulated samples. A representative experiment ( a ) and quantification of recruitment of indicated proteins to TNF-RSC from three independent experiments normalized to WT cells, mean + SEM ( b ). c , d Immunoblot analysis of lysates of ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cells stimulated with TNF (100 ng/ml) as indicated. A representative experiment ( c ) and quantification of three independent experiments normalized to the maximal response of WT cells, mean ± SEM ( d ). Statistical analysis was based on the area under the curve (AUC) for particular cell lines in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. e , f Immunoblot analysis of lysates from ST2 WT or Tank/Azi2 DKO cells that were unstimulated or stimulated for 24 h with TNF (250 ng/ml) and zVAD (20 µM) as indicated and subjected to RIPK1 immunoprecipitation. A representative experiment ( e ) and quantification of phosphorylated RIPK1 and cleaved Caspase-8 from three independent experiments normalized to WT cells stimulated with TNF alone, mean + SEM ( f ). g ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml), and the induction of cell death was monitored every 2 h using the <t>Incucyte</t> imaging system. The mean ± SEM from six separate experiments using two different Tank/Azi2 DKO ST2 clones is shown. Statistical analysis was based on the AUC for different cell lines and treatments in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. h ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml) alone or in the presence of zVAD (20 µM), Nec-1s (20 µM), or both inhibitors. Induction of cell death after 48 h was measured using the Incucyte imaging system. The mean + SEM from six independent experiments using two different Tank/Azi2 DKO ST2 clones is shown, two-way ANOVA with Tukey’s post-tests. n.s., not significant. #, nonspecific band.
Rapid Analyte Measurement Platform (Rampt) Wnv Test, supplied by Response Biomedical, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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par0 brandtdm controls  (Thermo Fisher)
90
Thermo Fisher par0 brandtdm controls
a , b Immunoblot analysis of TNF-RSC isolated from ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cell lines. Cells were stimulated with SF-TNF (500 ng/ml) as indicated, and lysates were subjected to anti-Flag immunoprecipitation. As a control, SF-TNF was added post-lysis to unstimulated samples. A representative experiment ( a ) and quantification of recruitment of indicated proteins to TNF-RSC from three independent experiments normalized to WT cells, mean + SEM ( b ). c , d Immunoblot analysis of lysates of ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cells stimulated with TNF (100 ng/ml) as indicated. A representative experiment ( c ) and quantification of three independent experiments normalized to the maximal response of WT cells, mean ± SEM ( d ). Statistical analysis was based on the area under the curve (AUC) for particular cell lines in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. e , f Immunoblot analysis of lysates from ST2 WT or Tank/Azi2 DKO cells that were unstimulated or stimulated for 24 h with TNF (250 ng/ml) and zVAD (20 µM) as indicated and subjected to RIPK1 immunoprecipitation. A representative experiment ( e ) and quantification of phosphorylated RIPK1 and cleaved Caspase-8 from three independent experiments normalized to WT cells stimulated with TNF alone, mean + SEM ( f ). g ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml), and the induction of cell death was monitored every 2 h using the <t>Incucyte</t> imaging system. The mean ± SEM from six separate experiments using two different Tank/Azi2 DKO ST2 clones is shown. Statistical analysis was based on the AUC for different cell lines and treatments in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. h ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml) alone or in the presence of zVAD (20 µM), Nec-1s (20 µM), or both inhibitors. Induction of cell death after 48 h was measured using the Incucyte imaging system. The mean + SEM from six independent experiments using two different Tank/Azi2 DKO ST2 clones is shown, two-way ANOVA with Tukey’s post-tests. n.s., not significant. #, nonspecific band.
Par0 Brandtdm Controls, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dade monitrol h controls  (Thermo Fisher)
90
Thermo Fisher dade monitrol h controls
a , b Immunoblot analysis of TNF-RSC isolated from ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cell lines. Cells were stimulated with SF-TNF (500 ng/ml) as indicated, and lysates were subjected to anti-Flag immunoprecipitation. As a control, SF-TNF was added post-lysis to unstimulated samples. A representative experiment ( a ) and quantification of recruitment of indicated proteins to TNF-RSC from three independent experiments normalized to WT cells, mean + SEM ( b ). c , d Immunoblot analysis of lysates of ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cells stimulated with TNF (100 ng/ml) as indicated. A representative experiment ( c ) and quantification of three independent experiments normalized to the maximal response of WT cells, mean ± SEM ( d ). Statistical analysis was based on the area under the curve (AUC) for particular cell lines in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. e , f Immunoblot analysis of lysates from ST2 WT or Tank/Azi2 DKO cells that were unstimulated or stimulated for 24 h with TNF (250 ng/ml) and zVAD (20 µM) as indicated and subjected to RIPK1 immunoprecipitation. A representative experiment ( e ) and quantification of phosphorylated RIPK1 and cleaved Caspase-8 from three independent experiments normalized to WT cells stimulated with TNF alone, mean + SEM ( f ). g ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml), and the induction of cell death was monitored every 2 h using the <t>Incucyte</t> imaging system. The mean ± SEM from six separate experiments using two different Tank/Azi2 DKO ST2 clones is shown. Statistical analysis was based on the AUC for different cell lines and treatments in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. h ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml) alone or in the presence of zVAD (20 µM), Nec-1s (20 µM), or both inhibitors. Induction of cell death after 48 h was measured using the Incucyte imaging system. The mean + SEM from six independent experiments using two different Tank/Azi2 DKO ST2 clones is shown, two-way ANOVA with Tukey’s post-tests. n.s., not significant. #, nonspecific band.
Dade Monitrol H Controls, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


a , b Immunoblot analysis of TNF-RSC isolated from ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cell lines. Cells were stimulated with SF-TNF (500 ng/ml) as indicated, and lysates were subjected to anti-Flag immunoprecipitation. As a control, SF-TNF was added post-lysis to unstimulated samples. A representative experiment ( a ) and quantification of recruitment of indicated proteins to TNF-RSC from three independent experiments normalized to WT cells, mean + SEM ( b ). c , d Immunoblot analysis of lysates of ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cells stimulated with TNF (100 ng/ml) as indicated. A representative experiment ( c ) and quantification of three independent experiments normalized to the maximal response of WT cells, mean ± SEM ( d ). Statistical analysis was based on the area under the curve (AUC) for particular cell lines in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. e , f Immunoblot analysis of lysates from ST2 WT or Tank/Azi2 DKO cells that were unstimulated or stimulated for 24 h with TNF (250 ng/ml) and zVAD (20 µM) as indicated and subjected to RIPK1 immunoprecipitation. A representative experiment ( e ) and quantification of phosphorylated RIPK1 and cleaved Caspase-8 from three independent experiments normalized to WT cells stimulated with TNF alone, mean + SEM ( f ). g ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml), and the induction of cell death was monitored every 2 h using the Incucyte imaging system. The mean ± SEM from six separate experiments using two different Tank/Azi2 DKO ST2 clones is shown. Statistical analysis was based on the AUC for different cell lines and treatments in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. h ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml) alone or in the presence of zVAD (20 µM), Nec-1s (20 µM), or both inhibitors. Induction of cell death after 48 h was measured using the Incucyte imaging system. The mean + SEM from six independent experiments using two different Tank/Azi2 DKO ST2 clones is shown, two-way ANOVA with Tukey’s post-tests. n.s., not significant. #, nonspecific band.

Journal: Nature Communications

Article Title: TBK1-associated adapters TANK and AZI2 protect mice against TNF-induced cell death and severe autoinflammatory diseases

doi: 10.1038/s41467-024-54399-4

Figure Lengend Snippet: a , b Immunoblot analysis of TNF-RSC isolated from ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cell lines. Cells were stimulated with SF-TNF (500 ng/ml) as indicated, and lysates were subjected to anti-Flag immunoprecipitation. As a control, SF-TNF was added post-lysis to unstimulated samples. A representative experiment ( a ) and quantification of recruitment of indicated proteins to TNF-RSC from three independent experiments normalized to WT cells, mean + SEM ( b ). c , d Immunoblot analysis of lysates of ST2 WT, Tank KO , Azi2 KO , or Tank/Azi2 DKO cells stimulated with TNF (100 ng/ml) as indicated. A representative experiment ( c ) and quantification of three independent experiments normalized to the maximal response of WT cells, mean ± SEM ( d ). Statistical analysis was based on the area under the curve (AUC) for particular cell lines in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. e , f Immunoblot analysis of lysates from ST2 WT or Tank/Azi2 DKO cells that were unstimulated or stimulated for 24 h with TNF (250 ng/ml) and zVAD (20 µM) as indicated and subjected to RIPK1 immunoprecipitation. A representative experiment ( e ) and quantification of phosphorylated RIPK1 and cleaved Caspase-8 from three independent experiments normalized to WT cells stimulated with TNF alone, mean + SEM ( f ). g ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml), and the induction of cell death was monitored every 2 h using the Incucyte imaging system. The mean ± SEM from six separate experiments using two different Tank/Azi2 DKO ST2 clones is shown. Statistical analysis was based on the AUC for different cell lines and treatments in each experiment, one-way ANOVA ( p -value shown) with Tukey´s post-tests. h ST2 WT and Tank/Azi2 DKO cells were unstimulated or stimulated with TNF (100 ng/ml) alone or in the presence of zVAD (20 µM), Nec-1s (20 µM), or both inhibitors. Induction of cell death after 48 h was measured using the Incucyte imaging system. The mean + SEM from six independent experiments using two different Tank/Azi2 DKO ST2 clones is shown, two-way ANOVA with Tukey’s post-tests. n.s., not significant. #, nonspecific band.

Article Snippet: The induction of cell death was assessed at two-hour intervals using Incucyte Live-Cell Analysis Systems (Sartorius).

Techniques: Western Blot, Isolation, Immunoprecipitation, Control, Lysis, Imaging, Clone Assay

a , b Immunoblot analysis of lysates of bone marrow-derived macrophages (BMDMs) isolated from WT or Tank/Azi2 DKO mice and stimulated with TNF (100 ng/ml) as indicated. A representative experiment ( a ) and quantification of three independent experiments using BMDMs independently isolated from 3 animal pairs. Data were normalized to the maximal response of WT cells, mean ± SEM ( b ). Statistical analysis was based on the AUC for each cell line, unpaired two-tailed t-test. c , d RNAseq analysis of WT and Tank/Azi2 DKO BMDMs stimulated for two hours with TNF (100 ng/ml). The BMDMs were independently isolated from 3 pairs of animals, and RNA was isolated in five separate experiments. Heatmap shows the expression of selected TNF-responsive genes ( c ). Gene set enrichment analysis (GSEA) shows the enrichment of 105 genes induced by TNF stimulation (log 2 fold change > 1 and padj < 0.01; samples from WT and Tank/Azi2 DKO BMDMs were analyzed together). The ranked gene list represents the contrast between unstimulated Tank/Azi2 DKO and WT BMDMs (blue) or stimulated Tank/Azi2 DKO and WT BMDMs (red). The p -value was estimated using an adaptive multi-level split Monte-Carlo scheme in the fgsea R package ( d ). e Analysis of cell death induced upon TNF (100 ng/ml) stimulation of WT or Tank/Azi2 DKO BMDMs. Induction of cell death was detected via the Incucyte imaging system. Mean + SEM from eight separate experiments using samples isolated from 4 animal pairs, two-way ANOVA with Tukey’s post-tests. n.s., not significant. #, nonspecific band.

Journal: Nature Communications

Article Title: TBK1-associated adapters TANK and AZI2 protect mice against TNF-induced cell death and severe autoinflammatory diseases

doi: 10.1038/s41467-024-54399-4

Figure Lengend Snippet: a , b Immunoblot analysis of lysates of bone marrow-derived macrophages (BMDMs) isolated from WT or Tank/Azi2 DKO mice and stimulated with TNF (100 ng/ml) as indicated. A representative experiment ( a ) and quantification of three independent experiments using BMDMs independently isolated from 3 animal pairs. Data were normalized to the maximal response of WT cells, mean ± SEM ( b ). Statistical analysis was based on the AUC for each cell line, unpaired two-tailed t-test. c , d RNAseq analysis of WT and Tank/Azi2 DKO BMDMs stimulated for two hours with TNF (100 ng/ml). The BMDMs were independently isolated from 3 pairs of animals, and RNA was isolated in five separate experiments. Heatmap shows the expression of selected TNF-responsive genes ( c ). Gene set enrichment analysis (GSEA) shows the enrichment of 105 genes induced by TNF stimulation (log 2 fold change > 1 and padj < 0.01; samples from WT and Tank/Azi2 DKO BMDMs were analyzed together). The ranked gene list represents the contrast between unstimulated Tank/Azi2 DKO and WT BMDMs (blue) or stimulated Tank/Azi2 DKO and WT BMDMs (red). The p -value was estimated using an adaptive multi-level split Monte-Carlo scheme in the fgsea R package ( d ). e Analysis of cell death induced upon TNF (100 ng/ml) stimulation of WT or Tank/Azi2 DKO BMDMs. Induction of cell death was detected via the Incucyte imaging system. Mean + SEM from eight separate experiments using samples isolated from 4 animal pairs, two-way ANOVA with Tukey’s post-tests. n.s., not significant. #, nonspecific band.

Article Snippet: The induction of cell death was assessed at two-hour intervals using Incucyte Live-Cell Analysis Systems (Sartorius).

Techniques: Western Blot, Derivative Assay, Isolation, Two Tailed Test, Expressing, Imaging