Cell Culture:Article Title: TGFβ-activated kinase-1 knockdown in hematopoietic stem-progenitor cells causes PANoptosis and myelodysplastic syndrome-like disease in mice.
Article Snippet: Primer names Primer sequences Product sizes TAK1 f1 5’-GGCTTTCATTGTGGAGGTAAGCTGAGA-3’ 320bp for floxed allele TAK1 r1 5’-GGAACCCGTGGATAAGTGCACTTGAAT-3’ 280bp for WT allele TAK1 f2 5’-GCAACTTCGACAACTTGCCTTCCTGTG-3’ TAK1 r2 5’-GCACTTGAATTAGCGGCCGCAAGCTTATAACT-3’ 1000bp for mutant allele Cre1 5'-CTAGGCCACAGAATTGAAAGATCT-3' Cre2 5'-GTAGGTGGAAATTCTAGCATCATC C-3' 324 bp for Cre Cre3 5'-GCGGTCTGGCAGTAAAAACTATC-3' Cre4 5'-GTGAAACAGCATTGCTGTCACTT-3' 100 bp for internal control Map3k7 Mm01191861_m1 ThermoFisher Scientific Catalog # 4351372 Gapdh Mm99999915_g1 ThermoFisher Scientific Catalog # 4331182 Table S2. .. Chemicals Venders Catalog No. Small molecular inhibitors Birinapant (xIAP inhibitor or Smac mimetic) Medchemexpress HY-16591 GNE684 (murine RIPK1 inhibitor) Medchemexpress HY-128585 GSK3145095 (Human RIPK1 inhibitor) Medchemexpress HY-111946 HS-276 (TAK1 inhibitor) Medchemexpress HY-147141E1155 Necrostatin 1S (Nec-1S) Medchemexpress HY-14622A VX-765 (Belnacasan) Caspase-1/4 inhibitor Selleckchem No.S2228 Z-VAD-FMK (Pan-caspase) Selleckchem No.S7023 Z-IETD-FMK (Caspase-8) Selleckchem S7314 CA-074 Me (cathepsin B) Medchemexpress HY-103350 Cell culture reagents Luciferase shRNA (shLuc)-tGFP lentiviral plasmid Origene TR30023 Map3k7 Mouse shRNA Plasmid (Locus ID 26409) Origene TL516837 1× Ham’s F-12 Nutrient Mix liquid media Life Technologies (Gibco) Insulin-Transferrin-Selenium-Ethanolamine (ITS-X, 100X) Thermo Fisher Scientific 51500-056 Penicillin-Streptomycin-L-Glutamine Solution (P/S/G, 100X) Thermo Fisher Scientific 10378016 N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) Thermo Fisher Scientific Gibco Polyvinyl alcohol (PVA) Sigma P8136 Mouse TPO (Thrombopoietin) Recombinant Protein, PeproTech® (rmTPO) Thermo Fisher Scientific Gibco 315-14 Mouse SCF Recombinant Protein, PeproTech® (rmSCF) Thermo Fisher ScientificGibco 250-03 IMDM (Iscove's Modified Dulbecco's Medium) Life Technologies (Gibco) 12440-053 100× Insulin-Transferrin-Selenium-ethanolamine (ITS-X) Life Technologies (Gibco) 51500-056 100× Penicillin-Streptomycin-Glutamine (P/S/G) 10378-016 10 740Y-P InvivoChem V2540 Butyzamide InvivoChem V51526 UM171 InvivoChem V1894 PCL-PVAc-PEG (Soluplus®) BASF Pharma 402932-23-4 Human PB plasma Stem Cell Technologies 70039.3 Human AB serum Sigma-Aldrich H4522 Holo-human transferrin Sigma-Aldrich T4132 heparin Stem Cell Technologies 7980 human insulin Sigma-Aldrich I9278 rhSCF PeproTech 300-07 rhIL-3 PeproTech 200-03 rhEPO PeproTech 100-64 MojoSortTM Mouse CD117 (c-Kit) Selection Kit Biolegend 480146 MethoCult® GF H4434 classic medium Stem Cell Technologies, Vancouver, BC 04434 MethoCult® GF M3434 medium Stem Cell Technologies, Vancouver, BC 03434 Reagents for immunofluorescence staining Citrate Buffer (pH 6.0), Concentrate ThermoFisher Scientific 005000 eBioscienceTM IHC Antigen Retrieval Solution - Low pH (10X) ThermoFisher Scientific 00-4955-58 eBioscienceTM IHC Antigen Retrieval Solution - High pH (10X) ThermoFisher Scientific 00-4956-58 Universal antigen retrieval buffer BiCell Scientific BCASUB SlowFadeTM Diamond Antifade Mountant with DAPI ThermoFisher Scientific S36964 10X Blocking Buffer abcom ab126587 Table S4. ..
Luciferase:Article Title: TGFβ-activated kinase-1 knockdown in hematopoietic stem-progenitor cells causes PANoptosis and myelodysplastic syndrome-like disease in mice.
Article Snippet: Primer names Primer sequences Product sizes TAK1 f1 5’-GGCTTTCATTGTGGAGGTAAGCTGAGA-3’ 320bp for floxed allele TAK1 r1 5’-GGAACCCGTGGATAAGTGCACTTGAAT-3’ 280bp for WT allele TAK1 f2 5’-GCAACTTCGACAACTTGCCTTCCTGTG-3’ TAK1 r2 5’-GCACTTGAATTAGCGGCCGCAAGCTTATAACT-3’ 1000bp for mutant allele Cre1 5'-CTAGGCCACAGAATTGAAAGATCT-3' Cre2 5'-GTAGGTGGAAATTCTAGCATCATC C-3' 324 bp for Cre Cre3 5'-GCGGTCTGGCAGTAAAAACTATC-3' Cre4 5'-GTGAAACAGCATTGCTGTCACTT-3' 100 bp for internal control Map3k7 Mm01191861_m1 ThermoFisher Scientific Catalog # 4351372 Gapdh Mm99999915_g1 ThermoFisher Scientific Catalog # 4331182 Table S2. .. Chemicals Venders Catalog No. Small molecular inhibitors Birinapant (xIAP inhibitor or Smac mimetic) Medchemexpress HY-16591 GNE684 (murine RIPK1 inhibitor) Medchemexpress HY-128585 GSK3145095 (Human RIPK1 inhibitor) Medchemexpress HY-111946 HS-276 (TAK1 inhibitor) Medchemexpress HY-147141E1155 Necrostatin 1S (Nec-1S) Medchemexpress HY-14622A VX-765 (Belnacasan) Caspase-1/4 inhibitor Selleckchem No.S2228 Z-VAD-FMK (Pan-caspase) Selleckchem No.S7023 Z-IETD-FMK (Caspase-8) Selleckchem S7314 CA-074 Me (cathepsin B) Medchemexpress HY-103350 Cell culture reagents Luciferase shRNA (shLuc)-tGFP lentiviral plasmid Origene TR30023 Map3k7 Mouse shRNA Plasmid (Locus ID 26409) Origene TL516837 1× Ham’s F-12 Nutrient Mix liquid media Life Technologies (Gibco) Insulin-Transferrin-Selenium-Ethanolamine (ITS-X, 100X) Thermo Fisher Scientific 51500-056 Penicillin-Streptomycin-L-Glutamine Solution (P/S/G, 100X) Thermo Fisher Scientific 10378016 N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) Thermo Fisher Scientific Gibco Polyvinyl alcohol (PVA) Sigma P8136 Mouse TPO (Thrombopoietin) Recombinant Protein, PeproTech® (rmTPO) Thermo Fisher Scientific Gibco 315-14 Mouse SCF Recombinant Protein, PeproTech® (rmSCF) Thermo Fisher ScientificGibco 250-03 IMDM (Iscove's Modified Dulbecco's Medium) Life Technologies (Gibco) 12440-053 100× Insulin-Transferrin-Selenium-ethanolamine (ITS-X) Life Technologies (Gibco) 51500-056 100× Penicillin-Streptomycin-Glutamine (P/S/G) 10378-016 10 740Y-P InvivoChem V2540 Butyzamide InvivoChem V51526 UM171 InvivoChem V1894 PCL-PVAc-PEG (Soluplus®) BASF Pharma 402932-23-4 Human PB plasma Stem Cell Technologies 70039.3 Human AB serum Sigma-Aldrich H4522 Holo-human transferrin Sigma-Aldrich T4132 heparin Stem Cell Technologies 7980 human insulin Sigma-Aldrich I9278 rhSCF PeproTech 300-07 rhIL-3 PeproTech 200-03 rhEPO PeproTech 100-64 MojoSortTM Mouse CD117 (c-Kit) Selection Kit Biolegend 480146 MethoCult® GF H4434 classic medium Stem Cell Technologies, Vancouver, BC 04434 MethoCult® GF M3434 medium Stem Cell Technologies, Vancouver, BC 03434 Reagents for immunofluorescence staining Citrate Buffer (pH 6.0), Concentrate ThermoFisher Scientific 005000 eBioscienceTM IHC Antigen Retrieval Solution - Low pH (10X) ThermoFisher Scientific 00-4955-58 eBioscienceTM IHC Antigen Retrieval Solution - High pH (10X) ThermoFisher Scientific 00-4956-58 Universal antigen retrieval buffer BiCell Scientific BCASUB SlowFadeTM Diamond Antifade Mountant with DAPI ThermoFisher Scientific S36964 10X Blocking Buffer abcom ab126587 Table S4. ..
shRNA:Article Title: TGFβ-activated kinase-1 knockdown in hematopoietic stem-progenitor cells causes PANoptosis and myelodysplastic syndrome-like disease in mice.
Article Snippet: Primer names Primer sequences Product sizes TAK1 f1 5’-GGCTTTCATTGTGGAGGTAAGCTGAGA-3’ 320bp for floxed allele TAK1 r1 5’-GGAACCCGTGGATAAGTGCACTTGAAT-3’ 280bp for WT allele TAK1 f2 5’-GCAACTTCGACAACTTGCCTTCCTGTG-3’ TAK1 r2 5’-GCACTTGAATTAGCGGCCGCAAGCTTATAACT-3’ 1000bp for mutant allele Cre1 5'-CTAGGCCACAGAATTGAAAGATCT-3' Cre2 5'-GTAGGTGGAAATTCTAGCATCATC C-3' 324 bp for Cre Cre3 5'-GCGGTCTGGCAGTAAAAACTATC-3' Cre4 5'-GTGAAACAGCATTGCTGTCACTT-3' 100 bp for internal control Map3k7 Mm01191861_m1 ThermoFisher Scientific Catalog # 4351372 Gapdh Mm99999915_g1 ThermoFisher Scientific Catalog # 4331182 Table S2. .. Chemicals Venders Catalog No. Small molecular inhibitors Birinapant (xIAP inhibitor or Smac mimetic) Medchemexpress HY-16591 GNE684 (murine RIPK1 inhibitor) Medchemexpress HY-128585 GSK3145095 (Human RIPK1 inhibitor) Medchemexpress HY-111946 HS-276 (TAK1 inhibitor) Medchemexpress HY-147141E1155 Necrostatin 1S (Nec-1S) Medchemexpress HY-14622A VX-765 (Belnacasan) Caspase-1/4 inhibitor Selleckchem No.S2228 Z-VAD-FMK (Pan-caspase) Selleckchem No.S7023 Z-IETD-FMK (Caspase-8) Selleckchem S7314 CA-074 Me (cathepsin B) Medchemexpress HY-103350 Cell culture reagents Luciferase shRNA (shLuc)-tGFP lentiviral plasmid Origene TR30023 Map3k7 Mouse shRNA Plasmid (Locus ID 26409) Origene TL516837 1× Ham’s F-12 Nutrient Mix liquid media Life Technologies (Gibco) Insulin-Transferrin-Selenium-Ethanolamine (ITS-X, 100X) Thermo Fisher Scientific 51500-056 Penicillin-Streptomycin-L-Glutamine Solution (P/S/G, 100X) Thermo Fisher Scientific 10378016 N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) Thermo Fisher Scientific Gibco Polyvinyl alcohol (PVA) Sigma P8136 Mouse TPO (Thrombopoietin) Recombinant Protein, PeproTech® (rmTPO) Thermo Fisher Scientific Gibco 315-14 Mouse SCF Recombinant Protein, PeproTech® (rmSCF) Thermo Fisher ScientificGibco 250-03 IMDM (Iscove's Modified Dulbecco's Medium) Life Technologies (Gibco) 12440-053 100× Insulin-Transferrin-Selenium-ethanolamine (ITS-X) Life Technologies (Gibco) 51500-056 100× Penicillin-Streptomycin-Glutamine (P/S/G) 10378-016 10 740Y-P InvivoChem V2540 Butyzamide InvivoChem V51526 UM171 InvivoChem V1894 PCL-PVAc-PEG (Soluplus®) BASF Pharma 402932-23-4 Human PB plasma Stem Cell Technologies 70039.3 Human AB serum Sigma-Aldrich H4522 Holo-human transferrin Sigma-Aldrich T4132 heparin Stem Cell Technologies 7980 human insulin Sigma-Aldrich I9278 rhSCF PeproTech 300-07 rhIL-3 PeproTech 200-03 rhEPO PeproTech 100-64 MojoSortTM Mouse CD117 (c-Kit) Selection Kit Biolegend 480146 MethoCult® GF H4434 classic medium Stem Cell Technologies, Vancouver, BC 04434 MethoCult® GF M3434 medium Stem Cell Technologies, Vancouver, BC 03434 Reagents for immunofluorescence staining Citrate Buffer (pH 6.0), Concentrate ThermoFisher Scientific 005000 eBioscienceTM IHC Antigen Retrieval Solution - Low pH (10X) ThermoFisher Scientific 00-4955-58 eBioscienceTM IHC Antigen Retrieval Solution - High pH (10X) ThermoFisher Scientific 00-4956-58 Universal antigen retrieval buffer BiCell Scientific BCASUB SlowFadeTM Diamond Antifade Mountant with DAPI ThermoFisher Scientific S36964 10X Blocking Buffer abcom ab126587 Table S4. ..
Plasmid Preparation:Article Title: TGFβ-activated kinase-1 knockdown in hematopoietic stem-progenitor cells causes PANoptosis and myelodysplastic syndrome-like disease in mice.
Article Snippet: Primer names Primer sequences Product sizes TAK1 f1 5’-GGCTTTCATTGTGGAGGTAAGCTGAGA-3’ 320bp for floxed allele TAK1 r1 5’-GGAACCCGTGGATAAGTGCACTTGAAT-3’ 280bp for WT allele TAK1 f2 5’-GCAACTTCGACAACTTGCCTTCCTGTG-3’ TAK1 r2 5’-GCACTTGAATTAGCGGCCGCAAGCTTATAACT-3’ 1000bp for mutant allele Cre1 5'-CTAGGCCACAGAATTGAAAGATCT-3' Cre2 5'-GTAGGTGGAAATTCTAGCATCATC C-3' 324 bp for Cre Cre3 5'-GCGGTCTGGCAGTAAAAACTATC-3' Cre4 5'-GTGAAACAGCATTGCTGTCACTT-3' 100 bp for internal control Map3k7 Mm01191861_m1 ThermoFisher Scientific Catalog # 4351372 Gapdh Mm99999915_g1 ThermoFisher Scientific Catalog # 4331182 Table S2. .. Chemicals Venders Catalog No. Small molecular inhibitors Birinapant (xIAP inhibitor or Smac mimetic) Medchemexpress HY-16591 GNE684 (murine RIPK1 inhibitor) Medchemexpress HY-128585 GSK3145095 (Human RIPK1 inhibitor) Medchemexpress HY-111946 HS-276 (TAK1 inhibitor) Medchemexpress HY-147141E1155 Necrostatin 1S (Nec-1S) Medchemexpress HY-14622A VX-765 (Belnacasan) Caspase-1/4 inhibitor Selleckchem No.S2228 Z-VAD-FMK (Pan-caspase) Selleckchem No.S7023 Z-IETD-FMK (Caspase-8) Selleckchem S7314 CA-074 Me (cathepsin B) Medchemexpress HY-103350 Cell culture reagents Luciferase shRNA (shLuc)-tGFP lentiviral plasmid Origene TR30023 Map3k7 Mouse shRNA Plasmid (Locus ID 26409) Origene TL516837 1× Ham’s F-12 Nutrient Mix liquid media Life Technologies (Gibco) Insulin-Transferrin-Selenium-Ethanolamine (ITS-X, 100X) Thermo Fisher Scientific 51500-056 Penicillin-Streptomycin-L-Glutamine Solution (P/S/G, 100X) Thermo Fisher Scientific 10378016 N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) Thermo Fisher Scientific Gibco Polyvinyl alcohol (PVA) Sigma P8136 Mouse TPO (Thrombopoietin) Recombinant Protein, PeproTech® (rmTPO) Thermo Fisher Scientific Gibco 315-14 Mouse SCF Recombinant Protein, PeproTech® (rmSCF) Thermo Fisher ScientificGibco 250-03 IMDM (Iscove's Modified Dulbecco's Medium) Life Technologies (Gibco) 12440-053 100× Insulin-Transferrin-Selenium-ethanolamine (ITS-X) Life Technologies (Gibco) 51500-056 100× Penicillin-Streptomycin-Glutamine (P/S/G) 10378-016 10 740Y-P InvivoChem V2540 Butyzamide InvivoChem V51526 UM171 InvivoChem V1894 PCL-PVAc-PEG (Soluplus®) BASF Pharma 402932-23-4 Human PB plasma Stem Cell Technologies 70039.3 Human AB serum Sigma-Aldrich H4522 Holo-human transferrin Sigma-Aldrich T4132 heparin Stem Cell Technologies 7980 human insulin Sigma-Aldrich I9278 rhSCF PeproTech 300-07 rhIL-3 PeproTech 200-03 rhEPO PeproTech 100-64 MojoSortTM Mouse CD117 (c-Kit) Selection Kit Biolegend 480146 MethoCult® GF H4434 classic medium Stem Cell Technologies, Vancouver, BC 04434 MethoCult® GF M3434 medium Stem Cell Technologies, Vancouver, BC 03434 Reagents for immunofluorescence staining Citrate Buffer (pH 6.0), Concentrate ThermoFisher Scientific 005000 eBioscienceTM IHC Antigen Retrieval Solution - Low pH (10X) ThermoFisher Scientific 00-4955-58 eBioscienceTM IHC Antigen Retrieval Solution - High pH (10X) ThermoFisher Scientific 00-4956-58 Universal antigen retrieval buffer BiCell Scientific BCASUB SlowFadeTM Diamond Antifade Mountant with DAPI ThermoFisher Scientific S36964 10X Blocking Buffer abcom ab126587 Table S4. ..
Recombinant:Article Title: TGFβ-activated kinase-1 knockdown in hematopoietic stem-progenitor cells causes PANoptosis and myelodysplastic syndrome-like disease in mice.
Article Snippet: Primer names Primer sequences Product sizes TAK1 f1 5’-GGCTTTCATTGTGGAGGTAAGCTGAGA-3’ 320bp for floxed allele TAK1 r1 5’-GGAACCCGTGGATAAGTGCACTTGAAT-3’ 280bp for WT allele TAK1 f2 5’-GCAACTTCGACAACTTGCCTTCCTGTG-3’ TAK1 r2 5’-GCACTTGAATTAGCGGCCGCAAGCTTATAACT-3’ 1000bp for mutant allele Cre1 5'-CTAGGCCACAGAATTGAAAGATCT-3' Cre2 5'-GTAGGTGGAAATTCTAGCATCATC C-3' 324 bp for Cre Cre3 5'-GCGGTCTGGCAGTAAAAACTATC-3' Cre4 5'-GTGAAACAGCATTGCTGTCACTT-3' 100 bp for internal control Map3k7 Mm01191861_m1 ThermoFisher Scientific Catalog # 4351372 Gapdh Mm99999915_g1 ThermoFisher Scientific Catalog # 4331182 Table S2. .. Chemicals Venders Catalog No. Small molecular inhibitors Birinapant (xIAP inhibitor or Smac mimetic) Medchemexpress HY-16591 GNE684 (murine RIPK1 inhibitor) Medchemexpress HY-128585 GSK3145095 (Human RIPK1 inhibitor) Medchemexpress HY-111946 HS-276 (TAK1 inhibitor) Medchemexpress HY-147141E1155 Necrostatin 1S (Nec-1S) Medchemexpress HY-14622A VX-765 (Belnacasan) Caspase-1/4 inhibitor Selleckchem No.S2228 Z-VAD-FMK (Pan-caspase) Selleckchem No.S7023 Z-IETD-FMK (Caspase-8) Selleckchem S7314 CA-074 Me (cathepsin B) Medchemexpress HY-103350 Cell culture reagents Luciferase shRNA (shLuc)-tGFP lentiviral plasmid Origene TR30023 Map3k7 Mouse shRNA Plasmid (Locus ID 26409) Origene TL516837 1× Ham’s F-12 Nutrient Mix liquid media Life Technologies (Gibco) Insulin-Transferrin-Selenium-Ethanolamine (ITS-X, 100X) Thermo Fisher Scientific 51500-056 Penicillin-Streptomycin-L-Glutamine Solution (P/S/G, 100X) Thermo Fisher Scientific 10378016 N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) Thermo Fisher Scientific Gibco Polyvinyl alcohol (PVA) Sigma P8136 Mouse TPO (Thrombopoietin) Recombinant Protein, PeproTech® (rmTPO) Thermo Fisher Scientific Gibco 315-14 Mouse SCF Recombinant Protein, PeproTech® (rmSCF) Thermo Fisher ScientificGibco 250-03 IMDM (Iscove's Modified Dulbecco's Medium) Life Technologies (Gibco) 12440-053 100× Insulin-Transferrin-Selenium-ethanolamine (ITS-X) Life Technologies (Gibco) 51500-056 100× Penicillin-Streptomycin-Glutamine (P/S/G) 10378-016 10 740Y-P InvivoChem V2540 Butyzamide InvivoChem V51526 UM171 InvivoChem V1894 PCL-PVAc-PEG (Soluplus®) BASF Pharma 402932-23-4 Human PB plasma Stem Cell Technologies 70039.3 Human AB serum Sigma-Aldrich H4522 Holo-human transferrin Sigma-Aldrich T4132 heparin Stem Cell Technologies 7980 human insulin Sigma-Aldrich I9278 rhSCF PeproTech 300-07 rhIL-3 PeproTech 200-03 rhEPO PeproTech 100-64 MojoSortTM Mouse CD117 (c-Kit) Selection Kit Biolegend 480146 MethoCult® GF H4434 classic medium Stem Cell Technologies, Vancouver, BC 04434 MethoCult® GF M3434 medium Stem Cell Technologies, Vancouver, BC 03434 Reagents for immunofluorescence staining Citrate Buffer (pH 6.0), Concentrate ThermoFisher Scientific 005000 eBioscienceTM IHC Antigen Retrieval Solution - Low pH (10X) ThermoFisher Scientific 00-4955-58 eBioscienceTM IHC Antigen Retrieval Solution - High pH (10X) ThermoFisher Scientific 00-4956-58 Universal antigen retrieval buffer BiCell Scientific BCASUB SlowFadeTM Diamond Antifade Mountant with DAPI ThermoFisher Scientific S36964 10X Blocking Buffer abcom ab126587 Table S4. ..
Modification:Article Title: TGFβ-activated kinase-1 knockdown in hematopoietic stem-progenitor cells causes PANoptosis and myelodysplastic syndrome-like disease in mice.
Article Snippet: Primer names Primer sequences Product sizes TAK1 f1 5’-GGCTTTCATTGTGGAGGTAAGCTGAGA-3’ 320bp for floxed allele TAK1 r1 5’-GGAACCCGTGGATAAGTGCACTTGAAT-3’ 280bp for WT allele TAK1 f2 5’-GCAACTTCGACAACTTGCCTTCCTGTG-3’ TAK1 r2 5’-GCACTTGAATTAGCGGCCGCAAGCTTATAACT-3’ 1000bp for mutant allele Cre1 5'-CTAGGCCACAGAATTGAAAGATCT-3' Cre2 5'-GTAGGTGGAAATTCTAGCATCATC C-3' 324 bp for Cre Cre3 5'-GCGGTCTGGCAGTAAAAACTATC-3' Cre4 5'-GTGAAACAGCATTGCTGTCACTT-3' 100 bp for internal control Map3k7 Mm01191861_m1 ThermoFisher Scientific Catalog # 4351372 Gapdh Mm99999915_g1 ThermoFisher Scientific Catalog # 4331182 Table S2. .. Chemicals Venders Catalog No. Small molecular inhibitors Birinapant (xIAP inhibitor or Smac mimetic) Medchemexpress HY-16591 GNE684 (murine RIPK1 inhibitor) Medchemexpress HY-128585 GSK3145095 (Human RIPK1 inhibitor) Medchemexpress HY-111946 HS-276 (TAK1 inhibitor) Medchemexpress HY-147141E1155 Necrostatin 1S (Nec-1S) Medchemexpress HY-14622A VX-765 (Belnacasan) Caspase-1/4 inhibitor Selleckchem No.S2228 Z-VAD-FMK (Pan-caspase) Selleckchem No.S7023 Z-IETD-FMK (Caspase-8) Selleckchem S7314 CA-074 Me (cathepsin B) Medchemexpress HY-103350 Cell culture reagents Luciferase shRNA (shLuc)-tGFP lentiviral plasmid Origene TR30023 Map3k7 Mouse shRNA Plasmid (Locus ID 26409) Origene TL516837 1× Ham’s F-12 Nutrient Mix liquid media Life Technologies (Gibco) Insulin-Transferrin-Selenium-Ethanolamine (ITS-X, 100X) Thermo Fisher Scientific 51500-056 Penicillin-Streptomycin-L-Glutamine Solution (P/S/G, 100X) Thermo Fisher Scientific 10378016 N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) Thermo Fisher Scientific Gibco Polyvinyl alcohol (PVA) Sigma P8136 Mouse TPO (Thrombopoietin) Recombinant Protein, PeproTech® (rmTPO) Thermo Fisher Scientific Gibco 315-14 Mouse SCF Recombinant Protein, PeproTech® (rmSCF) Thermo Fisher ScientificGibco 250-03 IMDM (Iscove's Modified Dulbecco's Medium) Life Technologies (Gibco) 12440-053 100× Insulin-Transferrin-Selenium-ethanolamine (ITS-X) Life Technologies (Gibco) 51500-056 100× Penicillin-Streptomycin-Glutamine (P/S/G) 10378-016 10 740Y-P InvivoChem V2540 Butyzamide InvivoChem V51526 UM171 InvivoChem V1894 PCL-PVAc-PEG (Soluplus®) BASF Pharma 402932-23-4 Human PB plasma Stem Cell Technologies 70039.3 Human AB serum Sigma-Aldrich H4522 Holo-human transferrin Sigma-Aldrich T4132 heparin Stem Cell Technologies 7980 human insulin Sigma-Aldrich I9278 rhSCF PeproTech 300-07 rhIL-3 PeproTech 200-03 rhEPO PeproTech 100-64 MojoSortTM Mouse CD117 (c-Kit) Selection Kit Biolegend 480146 MethoCult® GF H4434 classic medium Stem Cell Technologies, Vancouver, BC 04434 MethoCult® GF M3434 medium Stem Cell Technologies, Vancouver, BC 03434 Reagents for immunofluorescence staining Citrate Buffer (pH 6.0), Concentrate ThermoFisher Scientific 005000 eBioscienceTM IHC Antigen Retrieval Solution - Low pH (10X) ThermoFisher Scientific 00-4955-58 eBioscienceTM IHC Antigen Retrieval Solution - High pH (10X) ThermoFisher Scientific 00-4956-58 Universal antigen retrieval buffer BiCell Scientific BCASUB SlowFadeTM Diamond Antifade Mountant with DAPI ThermoFisher Scientific S36964 10X Blocking Buffer abcom ab126587 Table S4. ..
Clinical Proteomics:Article Title: TGFβ-activated kinase-1 knockdown in hematopoietic stem-progenitor cells causes PANoptosis and myelodysplastic syndrome-like disease in mice.
Article Snippet: Primer names Primer sequences Product sizes TAK1 f1 5’-GGCTTTCATTGTGGAGGTAAGCTGAGA-3’ 320bp for floxed allele TAK1 r1 5’-GGAACCCGTGGATAAGTGCACTTGAAT-3’ 280bp for WT allele TAK1 f2 5’-GCAACTTCGACAACTTGCCTTCCTGTG-3’ TAK1 r2 5’-GCACTTGAATTAGCGGCCGCAAGCTTATAACT-3’ 1000bp for mutant allele Cre1 5'-CTAGGCCACAGAATTGAAAGATCT-3' Cre2 5'-GTAGGTGGAAATTCTAGCATCATC C-3' 324 bp for Cre Cre3 5'-GCGGTCTGGCAGTAAAAACTATC-3' Cre4 5'-GTGAAACAGCATTGCTGTCACTT-3' 100 bp for internal control Map3k7 Mm01191861_m1 ThermoFisher Scientific Catalog # 4351372 Gapdh Mm99999915_g1 ThermoFisher Scientific Catalog # 4331182 Table S2. .. Chemicals Venders Catalog No. Small molecular inhibitors Birinapant (xIAP inhibitor or Smac mimetic) Medchemexpress HY-16591 GNE684 (murine RIPK1 inhibitor) Medchemexpress HY-128585 GSK3145095 (Human RIPK1 inhibitor) Medchemexpress HY-111946 HS-276 (TAK1 inhibitor) Medchemexpress HY-147141E1155 Necrostatin 1S (Nec-1S) Medchemexpress HY-14622A VX-765 (Belnacasan) Caspase-1/4 inhibitor Selleckchem No.S2228 Z-VAD-FMK (Pan-caspase) Selleckchem No.S7023 Z-IETD-FMK (Caspase-8) Selleckchem S7314 CA-074 Me (cathepsin B) Medchemexpress HY-103350 Cell culture reagents Luciferase shRNA (shLuc)-tGFP lentiviral plasmid Origene TR30023 Map3k7 Mouse shRNA Plasmid (Locus ID 26409) Origene TL516837 1× Ham’s F-12 Nutrient Mix liquid media Life Technologies (Gibco) Insulin-Transferrin-Selenium-Ethanolamine (ITS-X, 100X) Thermo Fisher Scientific 51500-056 Penicillin-Streptomycin-L-Glutamine Solution (P/S/G, 100X) Thermo Fisher Scientific 10378016 N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) Thermo Fisher Scientific Gibco Polyvinyl alcohol (PVA) Sigma P8136 Mouse TPO (Thrombopoietin) Recombinant Protein, PeproTech® (rmTPO) Thermo Fisher Scientific Gibco 315-14 Mouse SCF Recombinant Protein, PeproTech® (rmSCF) Thermo Fisher ScientificGibco 250-03 IMDM (Iscove's Modified Dulbecco's Medium) Life Technologies (Gibco) 12440-053 100× Insulin-Transferrin-Selenium-ethanolamine (ITS-X) Life Technologies (Gibco) 51500-056 100× Penicillin-Streptomycin-Glutamine (P/S/G) 10378-016 10 740Y-P InvivoChem V2540 Butyzamide InvivoChem V51526 UM171 InvivoChem V1894 PCL-PVAc-PEG (Soluplus®) BASF Pharma 402932-23-4 Human PB plasma Stem Cell Technologies 70039.3 Human AB serum Sigma-Aldrich H4522 Holo-human transferrin Sigma-Aldrich T4132 heparin Stem Cell Technologies 7980 human insulin Sigma-Aldrich I9278 rhSCF PeproTech 300-07 rhIL-3 PeproTech 200-03 rhEPO PeproTech 100-64 MojoSortTM Mouse CD117 (c-Kit) Selection Kit Biolegend 480146 MethoCult® GF H4434 classic medium Stem Cell Technologies, Vancouver, BC 04434 MethoCult® GF M3434 medium Stem Cell Technologies, Vancouver, BC 03434 Reagents for immunofluorescence staining Citrate Buffer (pH 6.0), Concentrate ThermoFisher Scientific 005000 eBioscienceTM IHC Antigen Retrieval Solution - Low pH (10X) ThermoFisher Scientific 00-4955-58 eBioscienceTM IHC Antigen Retrieval Solution - High pH (10X) ThermoFisher Scientific 00-4956-58 Universal antigen retrieval buffer BiCell Scientific BCASUB SlowFadeTM Diamond Antifade Mountant with DAPI ThermoFisher Scientific S36964 10X Blocking Buffer abcom ab126587 Table S4. ..
Selection:Article Title: TGFβ-activated kinase-1 knockdown in hematopoietic stem-progenitor cells causes PANoptosis and myelodysplastic syndrome-like disease in mice.
Article Snippet: Primer names Primer sequences Product sizes TAK1 f1 5’-GGCTTTCATTGTGGAGGTAAGCTGAGA-3’ 320bp for floxed allele TAK1 r1 5’-GGAACCCGTGGATAAGTGCACTTGAAT-3’ 280bp for WT allele TAK1 f2 5’-GCAACTTCGACAACTTGCCTTCCTGTG-3’ TAK1 r2 5’-GCACTTGAATTAGCGGCCGCAAGCTTATAACT-3’ 1000bp for mutant allele Cre1 5'-CTAGGCCACAGAATTGAAAGATCT-3' Cre2 5'-GTAGGTGGAAATTCTAGCATCATC C-3' 324 bp for Cre Cre3 5'-GCGGTCTGGCAGTAAAAACTATC-3' Cre4 5'-GTGAAACAGCATTGCTGTCACTT-3' 100 bp for internal control Map3k7 Mm01191861_m1 ThermoFisher Scientific Catalog # 4351372 Gapdh Mm99999915_g1 ThermoFisher Scientific Catalog # 4331182 Table S2. .. Chemicals Venders Catalog No. Small molecular inhibitors Birinapant (xIAP inhibitor or Smac mimetic) Medchemexpress HY-16591 GNE684 (murine RIPK1 inhibitor) Medchemexpress HY-128585 GSK3145095 (Human RIPK1 inhibitor) Medchemexpress HY-111946 HS-276 (TAK1 inhibitor) Medchemexpress HY-147141E1155 Necrostatin 1S (Nec-1S) Medchemexpress HY-14622A VX-765 (Belnacasan) Caspase-1/4 inhibitor Selleckchem No.S2228 Z-VAD-FMK (Pan-caspase) Selleckchem No.S7023 Z-IETD-FMK (Caspase-8) Selleckchem S7314 CA-074 Me (cathepsin B) Medchemexpress HY-103350 Cell culture reagents Luciferase shRNA (shLuc)-tGFP lentiviral plasmid Origene TR30023 Map3k7 Mouse shRNA Plasmid (Locus ID 26409) Origene TL516837 1× Ham’s F-12 Nutrient Mix liquid media Life Technologies (Gibco) Insulin-Transferrin-Selenium-Ethanolamine (ITS-X, 100X) Thermo Fisher Scientific 51500-056 Penicillin-Streptomycin-L-Glutamine Solution (P/S/G, 100X) Thermo Fisher Scientific 10378016 N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) Thermo Fisher Scientific Gibco Polyvinyl alcohol (PVA) Sigma P8136 Mouse TPO (Thrombopoietin) Recombinant Protein, PeproTech® (rmTPO) Thermo Fisher Scientific Gibco 315-14 Mouse SCF Recombinant Protein, PeproTech® (rmSCF) Thermo Fisher ScientificGibco 250-03 IMDM (Iscove's Modified Dulbecco's Medium) Life Technologies (Gibco) 12440-053 100× Insulin-Transferrin-Selenium-ethanolamine (ITS-X) Life Technologies (Gibco) 51500-056 100× Penicillin-Streptomycin-Glutamine (P/S/G) 10378-016 10 740Y-P InvivoChem V2540 Butyzamide InvivoChem V51526 UM171 InvivoChem V1894 PCL-PVAc-PEG (Soluplus®) BASF Pharma 402932-23-4 Human PB plasma Stem Cell Technologies 70039.3 Human AB serum Sigma-Aldrich H4522 Holo-human transferrin Sigma-Aldrich T4132 heparin Stem Cell Technologies 7980 human insulin Sigma-Aldrich I9278 rhSCF PeproTech 300-07 rhIL-3 PeproTech 200-03 rhEPO PeproTech 100-64 MojoSortTM Mouse CD117 (c-Kit) Selection Kit Biolegend 480146 MethoCult® GF H4434 classic medium Stem Cell Technologies, Vancouver, BC 04434 MethoCult® GF M3434 medium Stem Cell Technologies, Vancouver, BC 03434 Reagents for immunofluorescence staining Citrate Buffer (pH 6.0), Concentrate ThermoFisher Scientific 005000 eBioscienceTM IHC Antigen Retrieval Solution - Low pH (10X) ThermoFisher Scientific 00-4955-58 eBioscienceTM IHC Antigen Retrieval Solution - High pH (10X) ThermoFisher Scientific 00-4956-58 Universal antigen retrieval buffer BiCell Scientific BCASUB SlowFadeTM Diamond Antifade Mountant with DAPI ThermoFisher Scientific S36964 10X Blocking Buffer abcom ab126587 Table S4. ..
Immunofluorescence:Article Title: TGFβ-activated kinase-1 knockdown in hematopoietic stem-progenitor cells causes PANoptosis and myelodysplastic syndrome-like disease in mice.
Article Snippet: Primer names Primer sequences Product sizes TAK1 f1 5’-GGCTTTCATTGTGGAGGTAAGCTGAGA-3’ 320bp for floxed allele TAK1 r1 5’-GGAACCCGTGGATAAGTGCACTTGAAT-3’ 280bp for WT allele TAK1 f2 5’-GCAACTTCGACAACTTGCCTTCCTGTG-3’ TAK1 r2 5’-GCACTTGAATTAGCGGCCGCAAGCTTATAACT-3’ 1000bp for mutant allele Cre1 5'-CTAGGCCACAGAATTGAAAGATCT-3' Cre2 5'-GTAGGTGGAAATTCTAGCATCATC C-3' 324 bp for Cre Cre3 5'-GCGGTCTGGCAGTAAAAACTATC-3' Cre4 5'-GTGAAACAGCATTGCTGTCACTT-3' 100 bp for internal control Map3k7 Mm01191861_m1 ThermoFisher Scientific Catalog # 4351372 Gapdh Mm99999915_g1 ThermoFisher Scientific Catalog # 4331182 Table S2. .. Chemicals Venders Catalog No. Small molecular inhibitors Birinapant (xIAP inhibitor or Smac mimetic) Medchemexpress HY-16591 GNE684 (murine RIPK1 inhibitor) Medchemexpress HY-128585 GSK3145095 (Human RIPK1 inhibitor) Medchemexpress HY-111946 HS-276 (TAK1 inhibitor) Medchemexpress HY-147141E1155 Necrostatin 1S (Nec-1S) Medchemexpress HY-14622A VX-765 (Belnacasan) Caspase-1/4 inhibitor Selleckchem No.S2228 Z-VAD-FMK (Pan-caspase) Selleckchem No.S7023 Z-IETD-FMK (Caspase-8) Selleckchem S7314 CA-074 Me (cathepsin B) Medchemexpress HY-103350 Cell culture reagents Luciferase shRNA (shLuc)-tGFP lentiviral plasmid Origene TR30023 Map3k7 Mouse shRNA Plasmid (Locus ID 26409) Origene TL516837 1× Ham’s F-12 Nutrient Mix liquid media Life Technologies (Gibco) Insulin-Transferrin-Selenium-Ethanolamine (ITS-X, 100X) Thermo Fisher Scientific 51500-056 Penicillin-Streptomycin-L-Glutamine Solution (P/S/G, 100X) Thermo Fisher Scientific 10378016 N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) Thermo Fisher Scientific Gibco Polyvinyl alcohol (PVA) Sigma P8136 Mouse TPO (Thrombopoietin) Recombinant Protein, PeproTech® (rmTPO) Thermo Fisher Scientific Gibco 315-14 Mouse SCF Recombinant Protein, PeproTech® (rmSCF) Thermo Fisher ScientificGibco 250-03 IMDM (Iscove's Modified Dulbecco's Medium) Life Technologies (Gibco) 12440-053 100× Insulin-Transferrin-Selenium-ethanolamine (ITS-X) Life Technologies (Gibco) 51500-056 100× Penicillin-Streptomycin-Glutamine (P/S/G) 10378-016 10 740Y-P InvivoChem V2540 Butyzamide InvivoChem V51526 UM171 InvivoChem V1894 PCL-PVAc-PEG (Soluplus®) BASF Pharma 402932-23-4 Human PB plasma Stem Cell Technologies 70039.3 Human AB serum Sigma-Aldrich H4522 Holo-human transferrin Sigma-Aldrich T4132 heparin Stem Cell Technologies 7980 human insulin Sigma-Aldrich I9278 rhSCF PeproTech 300-07 rhIL-3 PeproTech 200-03 rhEPO PeproTech 100-64 MojoSortTM Mouse CD117 (c-Kit) Selection Kit Biolegend 480146 MethoCult® GF H4434 classic medium Stem Cell Technologies, Vancouver, BC 04434 MethoCult® GF M3434 medium Stem Cell Technologies, Vancouver, BC 03434 Reagents for immunofluorescence staining Citrate Buffer (pH 6.0), Concentrate ThermoFisher Scientific 005000 eBioscienceTM IHC Antigen Retrieval Solution - Low pH (10X) ThermoFisher Scientific 00-4955-58 eBioscienceTM IHC Antigen Retrieval Solution - High pH (10X) ThermoFisher Scientific 00-4956-58 Universal antigen retrieval buffer BiCell Scientific BCASUB SlowFadeTM Diamond Antifade Mountant with DAPI ThermoFisher Scientific S36964 10X Blocking Buffer abcom ab126587 Table S4. ..
Staining:Article Title: TGFβ-activated kinase-1 knockdown in hematopoietic stem-progenitor cells causes PANoptosis and myelodysplastic syndrome-like disease in mice.
Article Snippet: Primer names Primer sequences Product sizes TAK1 f1 5’-GGCTTTCATTGTGGAGGTAAGCTGAGA-3’ 320bp for floxed allele TAK1 r1 5’-GGAACCCGTGGATAAGTGCACTTGAAT-3’ 280bp for WT allele TAK1 f2 5’-GCAACTTCGACAACTTGCCTTCCTGTG-3’ TAK1 r2 5’-GCACTTGAATTAGCGGCCGCAAGCTTATAACT-3’ 1000bp for mutant allele Cre1 5'-CTAGGCCACAGAATTGAAAGATCT-3' Cre2 5'-GTAGGTGGAAATTCTAGCATCATC C-3' 324 bp for Cre Cre3 5'-GCGGTCTGGCAGTAAAAACTATC-3' Cre4 5'-GTGAAACAGCATTGCTGTCACTT-3' 100 bp for internal control Map3k7 Mm01191861_m1 ThermoFisher Scientific Catalog # 4351372 Gapdh Mm99999915_g1 ThermoFisher Scientific Catalog # 4331182 Table S2. .. Chemicals Venders Catalog No. Small molecular inhibitors Birinapant (xIAP inhibitor or Smac mimetic) Medchemexpress HY-16591 GNE684 (murine RIPK1 inhibitor) Medchemexpress HY-128585 GSK3145095 (Human RIPK1 inhibitor) Medchemexpress HY-111946 HS-276 (TAK1 inhibitor) Medchemexpress HY-147141E1155 Necrostatin 1S (Nec-1S) Medchemexpress HY-14622A VX-765 (Belnacasan) Caspase-1/4 inhibitor Selleckchem No.S2228 Z-VAD-FMK (Pan-caspase) Selleckchem No.S7023 Z-IETD-FMK (Caspase-8) Selleckchem S7314 CA-074 Me (cathepsin B) Medchemexpress HY-103350 Cell culture reagents Luciferase shRNA (shLuc)-tGFP lentiviral plasmid Origene TR30023 Map3k7 Mouse shRNA Plasmid (Locus ID 26409) Origene TL516837 1× Ham’s F-12 Nutrient Mix liquid media Life Technologies (Gibco) Insulin-Transferrin-Selenium-Ethanolamine (ITS-X, 100X) Thermo Fisher Scientific 51500-056 Penicillin-Streptomycin-L-Glutamine Solution (P/S/G, 100X) Thermo Fisher Scientific 10378016 N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) Thermo Fisher Scientific Gibco Polyvinyl alcohol (PVA) Sigma P8136 Mouse TPO (Thrombopoietin) Recombinant Protein, PeproTech® (rmTPO) Thermo Fisher Scientific Gibco 315-14 Mouse SCF Recombinant Protein, PeproTech® (rmSCF) Thermo Fisher ScientificGibco 250-03 IMDM (Iscove's Modified Dulbecco's Medium) Life Technologies (Gibco) 12440-053 100× Insulin-Transferrin-Selenium-ethanolamine (ITS-X) Life Technologies (Gibco) 51500-056 100× Penicillin-Streptomycin-Glutamine (P/S/G) 10378-016 10 740Y-P InvivoChem V2540 Butyzamide InvivoChem V51526 UM171 InvivoChem V1894 PCL-PVAc-PEG (Soluplus®) BASF Pharma 402932-23-4 Human PB plasma Stem Cell Technologies 70039.3 Human AB serum Sigma-Aldrich H4522 Holo-human transferrin Sigma-Aldrich T4132 heparin Stem Cell Technologies 7980 human insulin Sigma-Aldrich I9278 rhSCF PeproTech 300-07 rhIL-3 PeproTech 200-03 rhEPO PeproTech 100-64 MojoSortTM Mouse CD117 (c-Kit) Selection Kit Biolegend 480146 MethoCult® GF H4434 classic medium Stem Cell Technologies, Vancouver, BC 04434 MethoCult® GF M3434 medium Stem Cell Technologies, Vancouver, BC 03434 Reagents for immunofluorescence staining Citrate Buffer (pH 6.0), Concentrate ThermoFisher Scientific 005000 eBioscienceTM IHC Antigen Retrieval Solution - Low pH (10X) ThermoFisher Scientific 00-4955-58 eBioscienceTM IHC Antigen Retrieval Solution - High pH (10X) ThermoFisher Scientific 00-4956-58 Universal antigen retrieval buffer BiCell Scientific BCASUB SlowFadeTM Diamond Antifade Mountant with DAPI ThermoFisher Scientific S36964 10X Blocking Buffer abcom ab126587 Table S4. ..
Immunohistochemistry:Article Title: TGFβ-activated kinase-1 knockdown in hematopoietic stem-progenitor cells causes PANoptosis and myelodysplastic syndrome-like disease in mice.
Article Snippet: Primer names Primer sequences Product sizes TAK1 f1 5’-GGCTTTCATTGTGGAGGTAAGCTGAGA-3’ 320bp for floxed allele TAK1 r1 5’-GGAACCCGTGGATAAGTGCACTTGAAT-3’ 280bp for WT allele TAK1 f2 5’-GCAACTTCGACAACTTGCCTTCCTGTG-3’ TAK1 r2 5’-GCACTTGAATTAGCGGCCGCAAGCTTATAACT-3’ 1000bp for mutant allele Cre1 5'-CTAGGCCACAGAATTGAAAGATCT-3' Cre2 5'-GTAGGTGGAAATTCTAGCATCATC C-3' 324 bp for Cre Cre3 5'-GCGGTCTGGCAGTAAAAACTATC-3' Cre4 5'-GTGAAACAGCATTGCTGTCACTT-3' 100 bp for internal control Map3k7 Mm01191861_m1 ThermoFisher Scientific Catalog # 4351372 Gapdh Mm99999915_g1 ThermoFisher Scientific Catalog # 4331182 Table S2. .. Chemicals Venders Catalog No. Small molecular inhibitors Birinapant (xIAP inhibitor or Smac mimetic) Medchemexpress HY-16591 GNE684 (murine RIPK1 inhibitor) Medchemexpress HY-128585 GSK3145095 (Human RIPK1 inhibitor) Medchemexpress HY-111946 HS-276 (TAK1 inhibitor) Medchemexpress HY-147141E1155 Necrostatin 1S (Nec-1S) Medchemexpress HY-14622A VX-765 (Belnacasan) Caspase-1/4 inhibitor Selleckchem No.S2228 Z-VAD-FMK (Pan-caspase) Selleckchem No.S7023 Z-IETD-FMK (Caspase-8) Selleckchem S7314 CA-074 Me (cathepsin B) Medchemexpress HY-103350 Cell culture reagents Luciferase shRNA (shLuc)-tGFP lentiviral plasmid Origene TR30023 Map3k7 Mouse shRNA Plasmid (Locus ID 26409) Origene TL516837 1× Ham’s F-12 Nutrient Mix liquid media Life Technologies (Gibco) Insulin-Transferrin-Selenium-Ethanolamine (ITS-X, 100X) Thermo Fisher Scientific 51500-056 Penicillin-Streptomycin-L-Glutamine Solution (P/S/G, 100X) Thermo Fisher Scientific 10378016 N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) Thermo Fisher Scientific Gibco Polyvinyl alcohol (PVA) Sigma P8136 Mouse TPO (Thrombopoietin) Recombinant Protein, PeproTech® (rmTPO) Thermo Fisher Scientific Gibco 315-14 Mouse SCF Recombinant Protein, PeproTech® (rmSCF) Thermo Fisher ScientificGibco 250-03 IMDM (Iscove's Modified Dulbecco's Medium) Life Technologies (Gibco) 12440-053 100× Insulin-Transferrin-Selenium-ethanolamine (ITS-X) Life Technologies (Gibco) 51500-056 100× Penicillin-Streptomycin-Glutamine (P/S/G) 10378-016 10 740Y-P InvivoChem V2540 Butyzamide InvivoChem V51526 UM171 InvivoChem V1894 PCL-PVAc-PEG (Soluplus®) BASF Pharma 402932-23-4 Human PB plasma Stem Cell Technologies 70039.3 Human AB serum Sigma-Aldrich H4522 Holo-human transferrin Sigma-Aldrich T4132 heparin Stem Cell Technologies 7980 human insulin Sigma-Aldrich I9278 rhSCF PeproTech 300-07 rhIL-3 PeproTech 200-03 rhEPO PeproTech 100-64 MojoSortTM Mouse CD117 (c-Kit) Selection Kit Biolegend 480146 MethoCult® GF H4434 classic medium Stem Cell Technologies, Vancouver, BC 04434 MethoCult® GF M3434 medium Stem Cell Technologies, Vancouver, BC 03434 Reagents for immunofluorescence staining Citrate Buffer (pH 6.0), Concentrate ThermoFisher Scientific 005000 eBioscienceTM IHC Antigen Retrieval Solution - Low pH (10X) ThermoFisher Scientific 00-4955-58 eBioscienceTM IHC Antigen Retrieval Solution - High pH (10X) ThermoFisher Scientific 00-4956-58 Universal antigen retrieval buffer BiCell Scientific BCASUB SlowFadeTM Diamond Antifade Mountant with DAPI ThermoFisher Scientific S36964 10X Blocking Buffer abcom ab126587 Table S4. ..
Blocking Assay:Article Title: TGFβ-activated kinase-1 knockdown in hematopoietic stem-progenitor cells causes PANoptosis and myelodysplastic syndrome-like disease in mice.
Article Snippet: Primer names Primer sequences Product sizes TAK1 f1 5’-GGCTTTCATTGTGGAGGTAAGCTGAGA-3’ 320bp for floxed allele TAK1 r1 5’-GGAACCCGTGGATAAGTGCACTTGAAT-3’ 280bp for WT allele TAK1 f2 5’-GCAACTTCGACAACTTGCCTTCCTGTG-3’ TAK1 r2 5’-GCACTTGAATTAGCGGCCGCAAGCTTATAACT-3’ 1000bp for mutant allele Cre1 5'-CTAGGCCACAGAATTGAAAGATCT-3' Cre2 5'-GTAGGTGGAAATTCTAGCATCATC C-3' 324 bp for Cre Cre3 5'-GCGGTCTGGCAGTAAAAACTATC-3' Cre4 5'-GTGAAACAGCATTGCTGTCACTT-3' 100 bp for internal control Map3k7 Mm01191861_m1 ThermoFisher Scientific Catalog # 4351372 Gapdh Mm99999915_g1 ThermoFisher Scientific Catalog # 4331182 Table S2. .. Chemicals Venders Catalog No. Small molecular inhibitors Birinapant (xIAP inhibitor or Smac mimetic) Medchemexpress HY-16591 GNE684 (murine RIPK1 inhibitor) Medchemexpress HY-128585 GSK3145095 (Human RIPK1 inhibitor) Medchemexpress HY-111946 HS-276 (TAK1 inhibitor) Medchemexpress HY-147141E1155 Necrostatin 1S (Nec-1S) Medchemexpress HY-14622A VX-765 (Belnacasan) Caspase-1/4 inhibitor Selleckchem No.S2228 Z-VAD-FMK (Pan-caspase) Selleckchem No.S7023 Z-IETD-FMK (Caspase-8) Selleckchem S7314 CA-074 Me (cathepsin B) Medchemexpress HY-103350 Cell culture reagents Luciferase shRNA (shLuc)-tGFP lentiviral plasmid Origene TR30023 Map3k7 Mouse shRNA Plasmid (Locus ID 26409) Origene TL516837 1× Ham’s F-12 Nutrient Mix liquid media Life Technologies (Gibco) Insulin-Transferrin-Selenium-Ethanolamine (ITS-X, 100X) Thermo Fisher Scientific 51500-056 Penicillin-Streptomycin-L-Glutamine Solution (P/S/G, 100X) Thermo Fisher Scientific 10378016 N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) Thermo Fisher Scientific Gibco Polyvinyl alcohol (PVA) Sigma P8136 Mouse TPO (Thrombopoietin) Recombinant Protein, PeproTech® (rmTPO) Thermo Fisher Scientific Gibco 315-14 Mouse SCF Recombinant Protein, PeproTech® (rmSCF) Thermo Fisher ScientificGibco 250-03 IMDM (Iscove's Modified Dulbecco's Medium) Life Technologies (Gibco) 12440-053 100× Insulin-Transferrin-Selenium-ethanolamine (ITS-X) Life Technologies (Gibco) 51500-056 100× Penicillin-Streptomycin-Glutamine (P/S/G) 10378-016 10 740Y-P InvivoChem V2540 Butyzamide InvivoChem V51526 UM171 InvivoChem V1894 PCL-PVAc-PEG (Soluplus®) BASF Pharma 402932-23-4 Human PB plasma Stem Cell Technologies 70039.3 Human AB serum Sigma-Aldrich H4522 Holo-human transferrin Sigma-Aldrich T4132 heparin Stem Cell Technologies 7980 human insulin Sigma-Aldrich I9278 rhSCF PeproTech 300-07 rhIL-3 PeproTech 200-03 rhEPO PeproTech 100-64 MojoSortTM Mouse CD117 (c-Kit) Selection Kit Biolegend 480146 MethoCult® GF H4434 classic medium Stem Cell Technologies, Vancouver, BC 04434 MethoCult® GF M3434 medium Stem Cell Technologies, Vancouver, BC 03434 Reagents for immunofluorescence staining Citrate Buffer (pH 6.0), Concentrate ThermoFisher Scientific 005000 eBioscienceTM IHC Antigen Retrieval Solution - Low pH (10X) ThermoFisher Scientific 00-4955-58 eBioscienceTM IHC Antigen Retrieval Solution - High pH (10X) ThermoFisher Scientific 00-4956-58 Universal antigen retrieval buffer BiCell Scientific BCASUB SlowFadeTM Diamond Antifade Mountant with DAPI ThermoFisher Scientific S36964 10X Blocking Buffer abcom ab126587 Table S4. ..
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