laf237 Search Results


molecule inhibitors  (MedChemExpress)
93
MedChemExpress molecule inhibitors
A. DPP8/9 <t>inhibitors</t> cause ASC-GFP speck formation in the presence of NLRP1. 293T-ASC-GFP cells were transfected NLRP1 expressing plasmids at a ratio of 1 μg/5x10 5 cells. Transfected cells were treated with Talabostat (0.2 μM) and 1G244 (10 μM) for 16 hours before GFP imaging. Scale bar=20 μm. B. DPP8/9 inhibition does not activate the NLRP3 inflammasome or enhance a NLRP1 pyrin-domain (PYD) mutant, p. M77T. 293T-ASC-GFP cells were transfected and treated as in A. C. Talabostat leads to ASC oligomerization independently of DPP4. 293T-ASC-GFP cells were transfected and treated as in A. Cell pellets were lysed in 1xTBS buffer with 1% NP-40. Insoluble pellets were subjected to crosslinking with 1 mM DSS for 15 mins at 37 °C and solubilized in 1xTBS with 1% SDS. D. DPP8/9 inhibition by Talabostat causes NLRP1 self-oligomerization. 293T cells were transfected with the respective constructs at a ratio of 2 μg/5x10 5 cells and treated with Talabostat (2 μM) two days after transfection for 16 hours. E. Alanine mutations in the DPP9 catalytic triad dominantly activate the NLRP1-ASC inflammasome. 293T-ASC-GFP cells were co-transfected with NLRP1 and the respective DPP9 constructs and imaged two days after transfection. F. Opposing roles of wild-type DPP9 and S759A mutant in mediating ASC-GFP oligomerization in the presence of NLRP1. DSS crosslinking was performed as in C. G. Validation of CRISPR/Cas9-mediated deletion of DPP9 and subsequent knockdown of DPP8. Cells were harvested and lysed in 1xTBS buffer with 1% NP-40 4 days after siRNA transfection. H. DPP9 deletion activates the NLRP1 inflammasome with partial compensation by DPP8. 293T-ASC-GFP cells were treated with control or siRNAs against DPP8 for 3 days before NLRP1 transfection. Related to
Molecule Inhibitors, supplied by MedChemExpress, 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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dpp-4 inhibitors laf-237  (Novartis)
90
Novartis dpp-4 inhibitors laf-237
A. DPP8/9 <t>inhibitors</t> cause ASC-GFP speck formation in the presence of NLRP1. 293T-ASC-GFP cells were transfected NLRP1 expressing plasmids at a ratio of 1 μg/5x10 5 cells. Transfected cells were treated with Talabostat (0.2 μM) and 1G244 (10 μM) for 16 hours before GFP imaging. Scale bar=20 μm. B. DPP8/9 inhibition does not activate the NLRP3 inflammasome or enhance a NLRP1 pyrin-domain (PYD) mutant, p. M77T. 293T-ASC-GFP cells were transfected and treated as in A. C. Talabostat leads to ASC oligomerization independently of DPP4. 293T-ASC-GFP cells were transfected and treated as in A. Cell pellets were lysed in 1xTBS buffer with 1% NP-40. Insoluble pellets were subjected to crosslinking with 1 mM DSS for 15 mins at 37 °C and solubilized in 1xTBS with 1% SDS. D. DPP8/9 inhibition by Talabostat causes NLRP1 self-oligomerization. 293T cells were transfected with the respective constructs at a ratio of 2 μg/5x10 5 cells and treated with Talabostat (2 μM) two days after transfection for 16 hours. E. Alanine mutations in the DPP9 catalytic triad dominantly activate the NLRP1-ASC inflammasome. 293T-ASC-GFP cells were co-transfected with NLRP1 and the respective DPP9 constructs and imaged two days after transfection. F. Opposing roles of wild-type DPP9 and S759A mutant in mediating ASC-GFP oligomerization in the presence of NLRP1. DSS crosslinking was performed as in C. G. Validation of CRISPR/Cas9-mediated deletion of DPP9 and subsequent knockdown of DPP8. Cells were harvested and lysed in 1xTBS buffer with 1% NP-40 4 days after siRNA transfection. H. DPP9 deletion activates the NLRP1 inflammasome with partial compensation by DPP8. 293T-ASC-GFP cells were treated with control or siRNAs against DPP8 for 3 days before NLRP1 transfection. Related to
Dpp 4 Inhibitors Laf 237, supplied by Novartis, 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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dppiv inhibitor nvp-laf237  (GLSynthesis Inc)
90
GLSynthesis Inc dppiv inhibitor nvp-laf237
A. DPP8/9 <t>inhibitors</t> cause ASC-GFP speck formation in the presence of NLRP1. 293T-ASC-GFP cells were transfected NLRP1 expressing plasmids at a ratio of 1 μg/5x10 5 cells. Transfected cells were treated with Talabostat (0.2 μM) and 1G244 (10 μM) for 16 hours before GFP imaging. Scale bar=20 μm. B. DPP8/9 inhibition does not activate the NLRP3 inflammasome or enhance a NLRP1 pyrin-domain (PYD) mutant, p. M77T. 293T-ASC-GFP cells were transfected and treated as in A. C. Talabostat leads to ASC oligomerization independently of DPP4. 293T-ASC-GFP cells were transfected and treated as in A. Cell pellets were lysed in 1xTBS buffer with 1% NP-40. Insoluble pellets were subjected to crosslinking with 1 mM DSS for 15 mins at 37 °C and solubilized in 1xTBS with 1% SDS. D. DPP8/9 inhibition by Talabostat causes NLRP1 self-oligomerization. 293T cells were transfected with the respective constructs at a ratio of 2 μg/5x10 5 cells and treated with Talabostat (2 μM) two days after transfection for 16 hours. E. Alanine mutations in the DPP9 catalytic triad dominantly activate the NLRP1-ASC inflammasome. 293T-ASC-GFP cells were co-transfected with NLRP1 and the respective DPP9 constructs and imaged two days after transfection. F. Opposing roles of wild-type DPP9 and S759A mutant in mediating ASC-GFP oligomerization in the presence of NLRP1. DSS crosslinking was performed as in C. G. Validation of CRISPR/Cas9-mediated deletion of DPP9 and subsequent knockdown of DPP8. Cells were harvested and lysed in 1xTBS buffer with 1% NP-40 4 days after siRNA transfection. H. DPP9 deletion activates the NLRP1 inflammasome with partial compensation by DPP8. 293T-ASC-GFP cells were treated with control or siRNAs against DPP8 for 3 days before NLRP1 transfection. Related to
Dppiv Inhibitor Nvp Laf237, supplied by GLSynthesis 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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compounds mk-0431, laf-237 and nvp-dpp728  (Ranbaxy Pharmaceuticals Canada)
90
Ranbaxy Pharmaceuticals Canada compounds mk-0431, laf-237 and nvp-dpp728
A. DPP8/9 <t>inhibitors</t> cause ASC-GFP speck formation in the presence of NLRP1. 293T-ASC-GFP cells were transfected NLRP1 expressing plasmids at a ratio of 1 μg/5x10 5 cells. Transfected cells were treated with Talabostat (0.2 μM) and 1G244 (10 μM) for 16 hours before GFP imaging. Scale bar=20 μm. B. DPP8/9 inhibition does not activate the NLRP3 inflammasome or enhance a NLRP1 pyrin-domain (PYD) mutant, p. M77T. 293T-ASC-GFP cells were transfected and treated as in A. C. Talabostat leads to ASC oligomerization independently of DPP4. 293T-ASC-GFP cells were transfected and treated as in A. Cell pellets were lysed in 1xTBS buffer with 1% NP-40. Insoluble pellets were subjected to crosslinking with 1 mM DSS for 15 mins at 37 °C and solubilized in 1xTBS with 1% SDS. D. DPP8/9 inhibition by Talabostat causes NLRP1 self-oligomerization. 293T cells were transfected with the respective constructs at a ratio of 2 μg/5x10 5 cells and treated with Talabostat (2 μM) two days after transfection for 16 hours. E. Alanine mutations in the DPP9 catalytic triad dominantly activate the NLRP1-ASC inflammasome. 293T-ASC-GFP cells were co-transfected with NLRP1 and the respective DPP9 constructs and imaged two days after transfection. F. Opposing roles of wild-type DPP9 and S759A mutant in mediating ASC-GFP oligomerization in the presence of NLRP1. DSS crosslinking was performed as in C. G. Validation of CRISPR/Cas9-mediated deletion of DPP9 and subsequent knockdown of DPP8. Cells were harvested and lysed in 1xTBS buffer with 1% NP-40 4 days after siRNA transfection. H. DPP9 deletion activates the NLRP1 inflammasome with partial compensation by DPP8. 293T-ASC-GFP cells were treated with control or siRNAs against DPP8 for 3 days before NLRP1 transfection. Related to
Compounds Mk 0431, Laf 237 And Nvp Dpp728, supplied by Ranbaxy Pharmaceuticals Canada, 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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laf-237 axon 1631  (Axon Medchem LLC)
90
Axon Medchem LLC laf-237 axon 1631
A. DPP8/9 <t>inhibitors</t> cause ASC-GFP speck formation in the presence of NLRP1. 293T-ASC-GFP cells were transfected NLRP1 expressing plasmids at a ratio of 1 μg/5x10 5 cells. Transfected cells were treated with Talabostat (0.2 μM) and 1G244 (10 μM) for 16 hours before GFP imaging. Scale bar=20 μm. B. DPP8/9 inhibition does not activate the NLRP3 inflammasome or enhance a NLRP1 pyrin-domain (PYD) mutant, p. M77T. 293T-ASC-GFP cells were transfected and treated as in A. C. Talabostat leads to ASC oligomerization independently of DPP4. 293T-ASC-GFP cells were transfected and treated as in A. Cell pellets were lysed in 1xTBS buffer with 1% NP-40. Insoluble pellets were subjected to crosslinking with 1 mM DSS for 15 mins at 37 °C and solubilized in 1xTBS with 1% SDS. D. DPP8/9 inhibition by Talabostat causes NLRP1 self-oligomerization. 293T cells were transfected with the respective constructs at a ratio of 2 μg/5x10 5 cells and treated with Talabostat (2 μM) two days after transfection for 16 hours. E. Alanine mutations in the DPP9 catalytic triad dominantly activate the NLRP1-ASC inflammasome. 293T-ASC-GFP cells were co-transfected with NLRP1 and the respective DPP9 constructs and imaged two days after transfection. F. Opposing roles of wild-type DPP9 and S759A mutant in mediating ASC-GFP oligomerization in the presence of NLRP1. DSS crosslinking was performed as in C. G. Validation of CRISPR/Cas9-mediated deletion of DPP9 and subsequent knockdown of DPP8. Cells were harvested and lysed in 1xTBS buffer with 1% NP-40 4 days after siRNA transfection. H. DPP9 deletion activates the NLRP1 inflammasome with partial compensation by DPP8. 293T-ASC-GFP cells were treated with control or siRNAs against DPP8 for 3 days before NLRP1 transfection. Related to
Laf 237 Axon 1631, supplied by Axon Medchem LLC, 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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laf237  (Novo Nordisk)
90
Novo Nordisk laf237
A. DPP8/9 <t>inhibitors</t> cause ASC-GFP speck formation in the presence of NLRP1. 293T-ASC-GFP cells were transfected NLRP1 expressing plasmids at a ratio of 1 μg/5x10 5 cells. Transfected cells were treated with Talabostat (0.2 μM) and 1G244 (10 μM) for 16 hours before GFP imaging. Scale bar=20 μm. B. DPP8/9 inhibition does not activate the NLRP3 inflammasome or enhance a NLRP1 pyrin-domain (PYD) mutant, p. M77T. 293T-ASC-GFP cells were transfected and treated as in A. C. Talabostat leads to ASC oligomerization independently of DPP4. 293T-ASC-GFP cells were transfected and treated as in A. Cell pellets were lysed in 1xTBS buffer with 1% NP-40. Insoluble pellets were subjected to crosslinking with 1 mM DSS for 15 mins at 37 °C and solubilized in 1xTBS with 1% SDS. D. DPP8/9 inhibition by Talabostat causes NLRP1 self-oligomerization. 293T cells were transfected with the respective constructs at a ratio of 2 μg/5x10 5 cells and treated with Talabostat (2 μM) two days after transfection for 16 hours. E. Alanine mutations in the DPP9 catalytic triad dominantly activate the NLRP1-ASC inflammasome. 293T-ASC-GFP cells were co-transfected with NLRP1 and the respective DPP9 constructs and imaged two days after transfection. F. Opposing roles of wild-type DPP9 and S759A mutant in mediating ASC-GFP oligomerization in the presence of NLRP1. DSS crosslinking was performed as in C. G. Validation of CRISPR/Cas9-mediated deletion of DPP9 and subsequent knockdown of DPP8. Cells were harvested and lysed in 1xTBS buffer with 1% NP-40 4 days after siRNA transfection. H. DPP9 deletion activates the NLRP1 inflammasome with partial compensation by DPP8. 293T-ASC-GFP cells were treated with control or siRNAs against DPP8 for 3 days before NLRP1 transfection. Related to
Laf237, supplied by Novo Nordisk, 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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vildagliptin (laf237)  (Jiangsu Hansoh Pharmaceutical Co)
90
Jiangsu Hansoh Pharmaceutical Co vildagliptin (laf237)
A. DPP8/9 <t>inhibitors</t> cause ASC-GFP speck formation in the presence of NLRP1. 293T-ASC-GFP cells were transfected NLRP1 expressing plasmids at a ratio of 1 μg/5x10 5 cells. Transfected cells were treated with Talabostat (0.2 μM) and 1G244 (10 μM) for 16 hours before GFP imaging. Scale bar=20 μm. B. DPP8/9 inhibition does not activate the NLRP3 inflammasome or enhance a NLRP1 pyrin-domain (PYD) mutant, p. M77T. 293T-ASC-GFP cells were transfected and treated as in A. C. Talabostat leads to ASC oligomerization independently of DPP4. 293T-ASC-GFP cells were transfected and treated as in A. Cell pellets were lysed in 1xTBS buffer with 1% NP-40. Insoluble pellets were subjected to crosslinking with 1 mM DSS for 15 mins at 37 °C and solubilized in 1xTBS with 1% SDS. D. DPP8/9 inhibition by Talabostat causes NLRP1 self-oligomerization. 293T cells were transfected with the respective constructs at a ratio of 2 μg/5x10 5 cells and treated with Talabostat (2 μM) two days after transfection for 16 hours. E. Alanine mutations in the DPP9 catalytic triad dominantly activate the NLRP1-ASC inflammasome. 293T-ASC-GFP cells were co-transfected with NLRP1 and the respective DPP9 constructs and imaged two days after transfection. F. Opposing roles of wild-type DPP9 and S759A mutant in mediating ASC-GFP oligomerization in the presence of NLRP1. DSS crosslinking was performed as in C. G. Validation of CRISPR/Cas9-mediated deletion of DPP9 and subsequent knockdown of DPP8. Cells were harvested and lysed in 1xTBS buffer with 1% NP-40 4 days after siRNA transfection. H. DPP9 deletion activates the NLRP1 inflammasome with partial compensation by DPP8. 293T-ASC-GFP cells were treated with control or siRNAs against DPP8 for 3 days before NLRP1 transfection. Related to
Vildagliptin (Laf237), supplied by Jiangsu Hansoh Pharmaceutical Co, 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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dpp-iv inhibitor laf237  (AstraZeneca ltd)
90
AstraZeneca ltd dpp-iv inhibitor laf237
A. DPP8/9 <t>inhibitors</t> cause ASC-GFP speck formation in the presence of NLRP1. 293T-ASC-GFP cells were transfected NLRP1 expressing plasmids at a ratio of 1 μg/5x10 5 cells. Transfected cells were treated with Talabostat (0.2 μM) and 1G244 (10 μM) for 16 hours before GFP imaging. Scale bar=20 μm. B. DPP8/9 inhibition does not activate the NLRP3 inflammasome or enhance a NLRP1 pyrin-domain (PYD) mutant, p. M77T. 293T-ASC-GFP cells were transfected and treated as in A. C. Talabostat leads to ASC oligomerization independently of DPP4. 293T-ASC-GFP cells were transfected and treated as in A. Cell pellets were lysed in 1xTBS buffer with 1% NP-40. Insoluble pellets were subjected to crosslinking with 1 mM DSS for 15 mins at 37 °C and solubilized in 1xTBS with 1% SDS. D. DPP8/9 inhibition by Talabostat causes NLRP1 self-oligomerization. 293T cells were transfected with the respective constructs at a ratio of 2 μg/5x10 5 cells and treated with Talabostat (2 μM) two days after transfection for 16 hours. E. Alanine mutations in the DPP9 catalytic triad dominantly activate the NLRP1-ASC inflammasome. 293T-ASC-GFP cells were co-transfected with NLRP1 and the respective DPP9 constructs and imaged two days after transfection. F. Opposing roles of wild-type DPP9 and S759A mutant in mediating ASC-GFP oligomerization in the presence of NLRP1. DSS crosslinking was performed as in C. G. Validation of CRISPR/Cas9-mediated deletion of DPP9 and subsequent knockdown of DPP8. Cells were harvested and lysed in 1xTBS buffer with 1% NP-40 4 days after siRNA transfection. H. DPP9 deletion activates the NLRP1 inflammasome with partial compensation by DPP8. 293T-ASC-GFP cells were treated with control or siRNAs against DPP8 for 3 days before NLRP1 transfection. Related to
Dpp Iv Inhibitor Laf237, supplied by AstraZeneca ltd, 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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laf-237  (ApexBio)
90
ApexBio laf-237
A. DPP8/9 <t>inhibitors</t> cause ASC-GFP speck formation in the presence of NLRP1. 293T-ASC-GFP cells were transfected NLRP1 expressing plasmids at a ratio of 1 μg/5x10 5 cells. Transfected cells were treated with Talabostat (0.2 μM) and 1G244 (10 μM) for 16 hours before GFP imaging. Scale bar=20 μm. B. DPP8/9 inhibition does not activate the NLRP3 inflammasome or enhance a NLRP1 pyrin-domain (PYD) mutant, p. M77T. 293T-ASC-GFP cells were transfected and treated as in A. C. Talabostat leads to ASC oligomerization independently of DPP4. 293T-ASC-GFP cells were transfected and treated as in A. Cell pellets were lysed in 1xTBS buffer with 1% NP-40. Insoluble pellets were subjected to crosslinking with 1 mM DSS for 15 mins at 37 °C and solubilized in 1xTBS with 1% SDS. D. DPP8/9 inhibition by Talabostat causes NLRP1 self-oligomerization. 293T cells were transfected with the respective constructs at a ratio of 2 μg/5x10 5 cells and treated with Talabostat (2 μM) two days after transfection for 16 hours. E. Alanine mutations in the DPP9 catalytic triad dominantly activate the NLRP1-ASC inflammasome. 293T-ASC-GFP cells were co-transfected with NLRP1 and the respective DPP9 constructs and imaged two days after transfection. F. Opposing roles of wild-type DPP9 and S759A mutant in mediating ASC-GFP oligomerization in the presence of NLRP1. DSS crosslinking was performed as in C. G. Validation of CRISPR/Cas9-mediated deletion of DPP9 and subsequent knockdown of DPP8. Cells were harvested and lysed in 1xTBS buffer with 1% NP-40 4 days after siRNA transfection. H. DPP9 deletion activates the NLRP1 inflammasome with partial compensation by DPP8. 293T-ASC-GFP cells were treated with control or siRNAs against DPP8 for 3 days before NLRP1 transfection. Related to
Laf 237, supplied by ApexBio, 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. DPP8/9 inhibitors cause ASC-GFP speck formation in the presence of NLRP1. 293T-ASC-GFP cells were transfected NLRP1 expressing plasmids at a ratio of 1 μg/5x10 5 cells. Transfected cells were treated with Talabostat (0.2 μM) and 1G244 (10 μM) for 16 hours before GFP imaging. Scale bar=20 μm. B. DPP8/9 inhibition does not activate the NLRP3 inflammasome or enhance a NLRP1 pyrin-domain (PYD) mutant, p. M77T. 293T-ASC-GFP cells were transfected and treated as in A. C. Talabostat leads to ASC oligomerization independently of DPP4. 293T-ASC-GFP cells were transfected and treated as in A. Cell pellets were lysed in 1xTBS buffer with 1% NP-40. Insoluble pellets were subjected to crosslinking with 1 mM DSS for 15 mins at 37 °C and solubilized in 1xTBS with 1% SDS. D. DPP8/9 inhibition by Talabostat causes NLRP1 self-oligomerization. 293T cells were transfected with the respective constructs at a ratio of 2 μg/5x10 5 cells and treated with Talabostat (2 μM) two days after transfection for 16 hours. E. Alanine mutations in the DPP9 catalytic triad dominantly activate the NLRP1-ASC inflammasome. 293T-ASC-GFP cells were co-transfected with NLRP1 and the respective DPP9 constructs and imaged two days after transfection. F. Opposing roles of wild-type DPP9 and S759A mutant in mediating ASC-GFP oligomerization in the presence of NLRP1. DSS crosslinking was performed as in C. G. Validation of CRISPR/Cas9-mediated deletion of DPP9 and subsequent knockdown of DPP8. Cells were harvested and lysed in 1xTBS buffer with 1% NP-40 4 days after siRNA transfection. H. DPP9 deletion activates the NLRP1 inflammasome with partial compensation by DPP8. 293T-ASC-GFP cells were treated with control or siRNAs against DPP8 for 3 days before NLRP1 transfection. Related to

Journal: bioRxiv

Article Title: DPP9 is an endogenous and direct inhibitor of the NLRP1 inflammasome that guards against human auto-inflammatory diseases

doi: 10.1101/260919

Figure Lengend Snippet: A. DPP8/9 inhibitors cause ASC-GFP speck formation in the presence of NLRP1. 293T-ASC-GFP cells were transfected NLRP1 expressing plasmids at a ratio of 1 μg/5x10 5 cells. Transfected cells were treated with Talabostat (0.2 μM) and 1G244 (10 μM) for 16 hours before GFP imaging. Scale bar=20 μm. B. DPP8/9 inhibition does not activate the NLRP3 inflammasome or enhance a NLRP1 pyrin-domain (PYD) mutant, p. M77T. 293T-ASC-GFP cells were transfected and treated as in A. C. Talabostat leads to ASC oligomerization independently of DPP4. 293T-ASC-GFP cells were transfected and treated as in A. Cell pellets were lysed in 1xTBS buffer with 1% NP-40. Insoluble pellets were subjected to crosslinking with 1 mM DSS for 15 mins at 37 °C and solubilized in 1xTBS with 1% SDS. D. DPP8/9 inhibition by Talabostat causes NLRP1 self-oligomerization. 293T cells were transfected with the respective constructs at a ratio of 2 μg/5x10 5 cells and treated with Talabostat (2 μM) two days after transfection for 16 hours. E. Alanine mutations in the DPP9 catalytic triad dominantly activate the NLRP1-ASC inflammasome. 293T-ASC-GFP cells were co-transfected with NLRP1 and the respective DPP9 constructs and imaged two days after transfection. F. Opposing roles of wild-type DPP9 and S759A mutant in mediating ASC-GFP oligomerization in the presence of NLRP1. DSS crosslinking was performed as in C. G. Validation of CRISPR/Cas9-mediated deletion of DPP9 and subsequent knockdown of DPP8. Cells were harvested and lysed in 1xTBS buffer with 1% NP-40 4 days after siRNA transfection. H. DPP9 deletion activates the NLRP1 inflammasome with partial compensation by DPP8. 293T-ASC-GFP cells were treated with control or siRNAs against DPP8 for 3 days before NLRP1 transfection. Related to

Article Snippet: The small molecule inhibitors used are vildagliptin (MedChemExpress), saxagliptin (MedChemExpress), TC-E 5007 (Tocris), butabindide oxalate (MedChemExpress), Talabostat and (MedChemExpress), 1G244 (Santa Cruz Biotechnology), LPS (Ultrapure, Escherichia coli O111:B4, SigmaAldrich) and nigericin 10 uM (Invivogen, #tlrl-nig).

Techniques: Transfection, Expressing, Imaging, Inhibition, Mutagenesis, Construct, CRISPR

A. IL-1B secretion from primary human keratinocytes and PBMCs upon Talabostat (2 μM) treatment. B. Human keratinocytes secrete IL-1B upon DPP9 S759A expression. Human keratinocytes were transfected with the respective plasmids with a ratio of 0.5 μg/ 5 x10 5 cells. Conditioned media were harvested 24 hours post transfection. C. Cykokine/chemokine response of keratinocytes to Talabostat is highly similar to MSPC/FKLC patient-derived keratinocytes harboring gain-of-function NLRP1 mutations. Luminex array was performed on conditioned media of Talabostat-treated keratinocytes. Cytokines/chemokines that were also enriched in MSPC/FKLC patient-derived primary keratinocytes are shown in red. D. Cytokine/chemokine analysis of PBMCs treated with 2 μM Talabostat. Luminex array was performed on conditioned media of Talabostat-treated PBMCs isolated from three donors. P-values were calculated based on Student‘s t-tests after log transformation. E. DPP8/9 inhibition by Talabostat causes dose-dependent IL-1B processing. Cultured immortalized keratinocytes were treated with different 0.2 μM, 2 μM, 20 μM and 200 μM Talabostat for 24 hours. Conditioned media was concentrated 10 times for SDS-PAGE. F. DPP8/9 inhibition by S759A and S759P leads to IL-1B processing and secretion. Keratinocytes were transfected with the DPP9 expressing constructs. Conditioned media was harvested 24 hours post-transfection. G. DPP8/9 inhibition by Talabostat causes endogenous ASC oligomerization. DSS crosslinking was performed as in . H. DPP8/9 inhibition by Talabostat leads to IL-1B processing and secretion in PBMCs. Conditioned media from PBMC (Donor 3) was used for SDS-PAGE without prior concentration. I. NLRP1 and ASC knockdown abrogates Talabostat-induced ASC oligomerization and IL-1B processing. Immortalized keratinocytes were treated with 2 μM Talabostat for 24 hours three days after siRNA incubation. Conditioned media was concentrated 10 times before SDS-PAGE. J. Genetic requirement of NLRP1 , ASC and CASP1 , but not NLRP3 in the effect of DPP8/9 inhibition. Immortalized keratinocytes treated with siRNAs and DPP8/9 inhibitors as in J. Conditioned media was diluted 1:5 before IL-1B ELISA. K. CRISPR/Cas9-mediated deletion of NLRP1 and ASC blocks Talabostat-induced pyroptosis. L. Dissection of the genetic requirement for inflammasome activation upon DPP8/9 inhibition in mouse bone marrow derived macrophages (BMDMs). Murine BMDMs from the indicated genotypes were primed with LPS (200 ng/ml), then treated with the indicated concentrations of Talabostat for 24 hours. Related to

Journal: bioRxiv

Article Title: DPP9 is an endogenous and direct inhibitor of the NLRP1 inflammasome that guards against human auto-inflammatory diseases

doi: 10.1101/260919

Figure Lengend Snippet: A. IL-1B secretion from primary human keratinocytes and PBMCs upon Talabostat (2 μM) treatment. B. Human keratinocytes secrete IL-1B upon DPP9 S759A expression. Human keratinocytes were transfected with the respective plasmids with a ratio of 0.5 μg/ 5 x10 5 cells. Conditioned media were harvested 24 hours post transfection. C. Cykokine/chemokine response of keratinocytes to Talabostat is highly similar to MSPC/FKLC patient-derived keratinocytes harboring gain-of-function NLRP1 mutations. Luminex array was performed on conditioned media of Talabostat-treated keratinocytes. Cytokines/chemokines that were also enriched in MSPC/FKLC patient-derived primary keratinocytes are shown in red. D. Cytokine/chemokine analysis of PBMCs treated with 2 μM Talabostat. Luminex array was performed on conditioned media of Talabostat-treated PBMCs isolated from three donors. P-values were calculated based on Student‘s t-tests after log transformation. E. DPP8/9 inhibition by Talabostat causes dose-dependent IL-1B processing. Cultured immortalized keratinocytes were treated with different 0.2 μM, 2 μM, 20 μM and 200 μM Talabostat for 24 hours. Conditioned media was concentrated 10 times for SDS-PAGE. F. DPP8/9 inhibition by S759A and S759P leads to IL-1B processing and secretion. Keratinocytes were transfected with the DPP9 expressing constructs. Conditioned media was harvested 24 hours post-transfection. G. DPP8/9 inhibition by Talabostat causes endogenous ASC oligomerization. DSS crosslinking was performed as in . H. DPP8/9 inhibition by Talabostat leads to IL-1B processing and secretion in PBMCs. Conditioned media from PBMC (Donor 3) was used for SDS-PAGE without prior concentration. I. NLRP1 and ASC knockdown abrogates Talabostat-induced ASC oligomerization and IL-1B processing. Immortalized keratinocytes were treated with 2 μM Talabostat for 24 hours three days after siRNA incubation. Conditioned media was concentrated 10 times before SDS-PAGE. J. Genetic requirement of NLRP1 , ASC and CASP1 , but not NLRP3 in the effect of DPP8/9 inhibition. Immortalized keratinocytes treated with siRNAs and DPP8/9 inhibitors as in J. Conditioned media was diluted 1:5 before IL-1B ELISA. K. CRISPR/Cas9-mediated deletion of NLRP1 and ASC blocks Talabostat-induced pyroptosis. L. Dissection of the genetic requirement for inflammasome activation upon DPP8/9 inhibition in mouse bone marrow derived macrophages (BMDMs). Murine BMDMs from the indicated genotypes were primed with LPS (200 ng/ml), then treated with the indicated concentrations of Talabostat for 24 hours. Related to

Article Snippet: The small molecule inhibitors used are vildagliptin (MedChemExpress), saxagliptin (MedChemExpress), TC-E 5007 (Tocris), butabindide oxalate (MedChemExpress), Talabostat and (MedChemExpress), 1G244 (Santa Cruz Biotechnology), LPS (Ultrapure, Escherichia coli O111:B4, SigmaAldrich) and nigericin 10 uM (Invivogen, #tlrl-nig).

Techniques: Expressing, Transfection, Derivative Assay, Luminex, Isolation, Transformation Assay, Inhibition, Cell Culture, SDS Page, Construct, Concentration Assay, Incubation, Enzyme-linked Immunosorbent Assay, CRISPR, Dissection, Activation Assay

A. Talabostat induces IL-1B secretion either alone or in cooperation with LPSpriming in primary human PBMCs. B. Overlap between Talabostat dependent cytokine/chemokine signature and MSPC/FKLC patient-derived keratinocytes. C. Luminex analysis of Talabostat-treated LPS-prestimulated PBMCs. D. List of cytokines/chemokines that are induced by Talabostat in PBMCs. E. Lytic cell death of Talabostat-treated keratinocytes. F. Talabostat induces ASC speck formation in immortalized human keratinocytes. G. Talabostat causes significant leukocyte cell death. H. The effect on IL-1B secretion by a panel of peptidase/protease inhibitors on immortalized keratinocytes. Compounds with known IC50 for DPP9<100 nM are colored red. I. Lytic death in keratinocytes upon Talabostat exposure requires NLRP1 and ASC, but not NLRP3. J. Caspase-1 knockdown abrogates the effect of Talabostat on IL-1B processing.

Journal: bioRxiv

Article Title: DPP9 is an endogenous and direct inhibitor of the NLRP1 inflammasome that guards against human auto-inflammatory diseases

doi: 10.1101/260919

Figure Lengend Snippet: A. Talabostat induces IL-1B secretion either alone or in cooperation with LPSpriming in primary human PBMCs. B. Overlap between Talabostat dependent cytokine/chemokine signature and MSPC/FKLC patient-derived keratinocytes. C. Luminex analysis of Talabostat-treated LPS-prestimulated PBMCs. D. List of cytokines/chemokines that are induced by Talabostat in PBMCs. E. Lytic cell death of Talabostat-treated keratinocytes. F. Talabostat induces ASC speck formation in immortalized human keratinocytes. G. Talabostat causes significant leukocyte cell death. H. The effect on IL-1B secretion by a panel of peptidase/protease inhibitors on immortalized keratinocytes. Compounds with known IC50 for DPP9<100 nM are colored red. I. Lytic death in keratinocytes upon Talabostat exposure requires NLRP1 and ASC, but not NLRP3. J. Caspase-1 knockdown abrogates the effect of Talabostat on IL-1B processing.

Article Snippet: The small molecule inhibitors used are vildagliptin (MedChemExpress), saxagliptin (MedChemExpress), TC-E 5007 (Tocris), butabindide oxalate (MedChemExpress), Talabostat and (MedChemExpress), 1G244 (Santa Cruz Biotechnology), LPS (Ultrapure, Escherichia coli O111:B4, SigmaAldrich) and nigericin 10 uM (Invivogen, #tlrl-nig).

Techniques: Derivative Assay, Luminex