Aqueous Normal-phase Chromatography:Article Title: Myosin inhibitor reverses hypertrophic cardiomyopathy in genotypically diverse pediatric iPSC-cardiomyocytes to mirror variant correction
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies OCT3/4 antibody PE-conjugated BD Biosciences Cat# 561556; RRID:AB_10714783 Sox2 PerCP-Cy5.5- conjugated BD Biosciences Cat# 561506; RRID:AB_10646039 SSEA-4 V450-conjugated BD Biosciences Cat# 561156; RRID:AB_10896140 TRA-1-60 AF647-conjugated Biolegend Cat# 330605; RRID:AB_1227813 a-actinin antibody Abcam Cat# Ab9465; RRID:AB_307264 MLC2v antibody Abcam Cat# Ab79935; RRID:AB_1952220 Actin Antibody R&D Systems Cat# MAB93081; RRID:AB_3068544 ANP antibody R&D Systems Cat# AF3366; RRID:AB_2915938 TNNI3 antibody R&D Systems Cat# MAB68871; RRID:AB_3068546 MYH7 antibody R&D Systems Cat# MAB9096; RRID:AB_2915972 MYBPC3 antibody Abcam Cat# Ab108522; RRID:AB_10865856 GAPDH antibody Abcam Cat# Ab8245; RRID:AB_2107448 Anti-rabbit 555 secondary antibody ThermoFisher Cat# A-21429; RRID:AB_2535850 Anti-mouse 488 secondary antibody ThermoFisher Cat# A-21121; RRID:AB_2535764 Anti-rabbit 647 secondary antibody ThermoFisher Cat# A-32795; RRID:AB_2762835 Anti-mouse 647 secondary antibody ThermoFisher Cat# A-21240; RRID:AB_2535809 Anti-mouse IgG HRP conjugated R&D Systems Cat# HAF018; RRID: AB_573130 Anti-rabbit IgG HRP conjugated R&D Systems Cat# HAF008; RRID:AB_357235 Biological samples Patient fibroblasts or lymphoblasts Sickkids Heart Center Biobank https://theheartcentrebiobank.com/ Chemicals, peptides, and recombinant proteins Puromycin Sigma-Aldrich Cat# P8833 Fibronectin ThermoFisher Cat# 33016015 Gelatin Sigma-Aldrich Cat# G9391 Matrigel Corning Cat# 354230 Collagen Corning Cat# 354249 PSC Cardiomyocyte Differentiation Kit ThermoFisher Cat# A2921201 STEMdiff cardiomyocyte differentiation kit STEMCELL Technologies Cat# 05010 STEMdiff cardiomyocyte dissociation kit STEMCELL Technologies Cat# 05025 iCell cardiomyocyte maintenance medium FUJIFILM Cellular Dymamics Cat# M1003 RPMI ThermoFisher Cat# 11875-093 B-27 ThermoFisher Cat# 17504-044 StemPro-34 ThermoFisher Cat# 10639011 CHIR-99021 MedChemExpress Cat# HY-10182 cOmplete protease inhibitor cocktail Sigma-Aldrich Cat# 11836170001 Protein A-agarose beads Cell Signaling Technology Cat# 9863 Protein G agarose beads Cell Signaling Technology Cat# 37478 Pierce ECL Western Blotting Substrate ThermoFisher Cat# 32109 (Continued on next page) Cell Reports Medicine 5, 101520, May 21, 2024 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER MYK-461 MedChemExpress Cat# SAR439152 Aficamten Cayman Chemical Cat# 36304 Verapamil Sigma-Aldrich Cat# V45629 Metoprolol Sigma-Aldrich Cat# M1830000 Isoproterenol Cayman Chemical Cat# 15592 DMSO Sigma-Aldrich Cat# D2650 DAPI Sigma-Aldrich Cat# 09564 Fluo-4 calcium dye ThermoFisher Cat# F14201 Critical commercial assays Precision gRNA Synthesis kit ThermoFisher Cat# A29377 ddPCRTM Supermix Biorad Cat# 1863024 LipofectamineTM Stem Transfection Reagent ThermoFisher Cat# STEM00015 Myco Alert kit Lonza Cat# LT07-418 ATPase assay kit Abcam Cat# Ab234055 Deposited data Whole genome sequencing European Genome-Phenome Archive (EGA) https://ega-archive.org/; EGAS00001004929 Experimental models: Cell lines PGPC17 Hildebrandt et al.27 PGPC17 MYH7 V606M patient cell line This study 80 MYH7 R453C patient cell line This study 81 MYBPC3 G148R patient cell line This study 82 MYBPC3 P955fs TNNI3 A157V patient cell line This study 83 MYH7 V606M corrected cell line This study 80C MYH7 R453C corrected cell line This study 81C MYBPC3 G148R corrected cell line This study 82C MYBPC3 P955fs corrected cell line This study 83C MYBPC3 G148R edited line This study G148R MYBPC3 R502W edited line This study R502W MYBPC3 Comp. het. edited line This study Comp. het.
Article Title: Myosin inhibitor reverses hypertrophic cardiomyopathy in genotypically diverse pediatric iPSC-cardiomyocytes to mirror variant correction.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies OCT3/4 antibody PE-conjugated BD Biosciences Cat# 561556; RRID:AB_10714783 Sox2 PerCP-Cy5.5- conjugated BD Biosciences Cat# 561506; RRID:AB_10646039 SSEA-4 V450-conjugated BD Biosciences Cat# 561156; RRID:AB_10896140 TRA-1-60 AF647-conjugated Biolegend Cat# 330605; RRID:AB_1227813 a-actinin antibody Abcam Cat# Ab9465; RRID:AB_307264 MLC2v antibody Abcam Cat# Ab79935; RRID:AB_1952220 Actin Antibody R&D Systems Cat# MAB93081; RRID:AB_3068544 ANP antibody R&D Systems Cat# AF3366; RRID:AB_2915938 TNNI3 antibody R&D Systems Cat# MAB68871; RRID:AB_3068546 MYH7 antibody R&D Systems Cat# MAB9096; RRID:AB_2915972 MYBPC3 antibody Abcam Cat# Ab108522; RRID:AB_10865856 GAPDH antibody Abcam Cat# Ab8245; RRID:AB_2107448 Anti-rabbit 555 secondary antibody ThermoFisher Cat# A-21429; RRID:AB_2535850 Anti-mouse 488 secondary antibody ThermoFisher Cat# A-21121; RRID:AB_2535764 Anti-rabbit 647 secondary antibody ThermoFisher Cat# A-32795; RRID:AB_2762835 Anti-mouse 647 secondary antibody ThermoFisher Cat# A-21240; RRID:AB_2535809 Anti-mouse IgG HRP conjugated R&D Systems Cat# HAF018; RRID: AB_573130 Anti-rabbit IgG HRP conjugated R&D Systems Cat# HAF008; RRID:AB_357235 Biological samples Patient fibroblasts or lymphoblasts Sickkids Heart Center Biobank https://theheartcentrebiobank.com/ Chemicals, peptides, and recombinant proteins Puromycin Sigma-Aldrich Cat# P8833 Fibronectin ThermoFisher Cat# 33016015 Gelatin Sigma-Aldrich Cat# G9391 Matrigel Corning Cat# 354230 Collagen Corning Cat# 354249 PSC Cardiomyocyte Differentiation Kit ThermoFisher Cat# A2921201 STEMdiff cardiomyocyte differentiation kit STEMCELL Technologies Cat# 05010 STEMdiff cardiomyocyte dissociation kit STEMCELL Technologies Cat# 05025 iCell cardiomyocyte maintenance medium FUJIFILM Cellular Dymamics Cat# M1003 RPMI ThermoFisher Cat# 11875-093 B-27 ThermoFisher Cat# 17504-044 StemPro-34 ThermoFisher Cat# 10639011 CHIR-99021 MedChemExpress Cat# HY-10182 cOmplete protease inhibitor cocktail Sigma-Aldrich Cat# 11836170001 Protein A-agarose beads Cell Signaling Technology Cat# 9863 Protein G agarose beads Cell Signaling Technology Cat# 37478 Pierce ECL Western Blotting Substrate ThermoFisher Cat# 32109 (Continued on next page) Cell Reports Medicine 5, 101520, May 21, 2024 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER MYK-461 MedChemExpress Cat# SAR439152 Aficamten Cayman Chemical Cat# 36304 Verapamil Sigma-Aldrich Cat# V45629 Metoprolol Sigma-Aldrich Cat# M1830000 Isoproterenol Cayman Chemical Cat# 15592 DMSO Sigma-Aldrich Cat# D2650 DAPI Sigma-Aldrich Cat# 09564 Fluo-4 calcium dye ThermoFisher Cat# F14201 Critical commercial assays Precision gRNA Synthesis kit ThermoFisher Cat# A29377 ddPCRTM Supermix Biorad Cat# 1863024 LipofectamineTM Stem Transfection Reagent ThermoFisher Cat# STEM00015 Myco Alert kit Lonza Cat# LT07-418 ATPase assay kit Abcam Cat# Ab234055 Deposited data Whole genome sequencing European Genome-Phenome Archive (EGA) https://ega-archive.org/; EGAS00001004929 Experimental models: Cell lines PGPC17 Hildebrandt et al.27 PGPC17 MYH7 V606M patient cell line This study 80 MYH7 R453C patient cell line This study 81 MYBPC3 G148R patient cell line This study 82 MYBPC3 P955fs TNNI3 A157V patient cell line This study 83 MYH7 V606M corrected cell line This study 80C MYH7 R453C corrected cell line This study 81C MYBPC3 G148R corrected cell line This study 82C MYBPC3 P955fs corrected cell line This study 83C MYBPC3 G148R edited line This study G148R MYBPC3 R502W edited line This study R502W MYBPC3 Comp. het. edited line This study Comp. het.
Recombinant:Article Title: Myosin inhibitor reverses hypertrophic cardiomyopathy in genotypically diverse pediatric iPSC-cardiomyocytes to mirror variant correction
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies OCT3/4 antibody PE-conjugated BD Biosciences Cat# 561556; RRID:AB_10714783 Sox2 PerCP-Cy5.5- conjugated BD Biosciences Cat# 561506; RRID:AB_10646039 SSEA-4 V450-conjugated BD Biosciences Cat# 561156; RRID:AB_10896140 TRA-1-60 AF647-conjugated Biolegend Cat# 330605; RRID:AB_1227813 a-actinin antibody Abcam Cat# Ab9465; RRID:AB_307264 MLC2v antibody Abcam Cat# Ab79935; RRID:AB_1952220 Actin Antibody R&D Systems Cat# MAB93081; RRID:AB_3068544 ANP antibody R&D Systems Cat# AF3366; RRID:AB_2915938 TNNI3 antibody R&D Systems Cat# MAB68871; RRID:AB_3068546 MYH7 antibody R&D Systems Cat# MAB9096; RRID:AB_2915972 MYBPC3 antibody Abcam Cat# Ab108522; RRID:AB_10865856 GAPDH antibody Abcam Cat# Ab8245; RRID:AB_2107448 Anti-rabbit 555 secondary antibody ThermoFisher Cat# A-21429; RRID:AB_2535850 Anti-mouse 488 secondary antibody ThermoFisher Cat# A-21121; RRID:AB_2535764 Anti-rabbit 647 secondary antibody ThermoFisher Cat# A-32795; RRID:AB_2762835 Anti-mouse 647 secondary antibody ThermoFisher Cat# A-21240; RRID:AB_2535809 Anti-mouse IgG HRP conjugated R&D Systems Cat# HAF018; RRID: AB_573130 Anti-rabbit IgG HRP conjugated R&D Systems Cat# HAF008; RRID:AB_357235 Biological samples Patient fibroblasts or lymphoblasts Sickkids Heart Center Biobank https://theheartcentrebiobank.com/ Chemicals, peptides, and recombinant proteins Puromycin Sigma-Aldrich Cat# P8833 Fibronectin ThermoFisher Cat# 33016015 Gelatin Sigma-Aldrich Cat# G9391 Matrigel Corning Cat# 354230 Collagen Corning Cat# 354249 PSC Cardiomyocyte Differentiation Kit ThermoFisher Cat# A2921201 STEMdiff cardiomyocyte differentiation kit STEMCELL Technologies Cat# 05010 STEMdiff cardiomyocyte dissociation kit STEMCELL Technologies Cat# 05025 iCell cardiomyocyte maintenance medium FUJIFILM Cellular Dymamics Cat# M1003 RPMI ThermoFisher Cat# 11875-093 B-27 ThermoFisher Cat# 17504-044 StemPro-34 ThermoFisher Cat# 10639011 CHIR-99021 MedChemExpress Cat# HY-10182 cOmplete protease inhibitor cocktail Sigma-Aldrich Cat# 11836170001 Protein A-agarose beads Cell Signaling Technology Cat# 9863 Protein G agarose beads Cell Signaling Technology Cat# 37478 Pierce ECL Western Blotting Substrate ThermoFisher Cat# 32109 (Continued on next page) Cell Reports Medicine 5, 101520, May 21, 2024 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER MYK-461 MedChemExpress Cat# SAR439152 Aficamten Cayman Chemical Cat# 36304 Verapamil Sigma-Aldrich Cat# V45629 Metoprolol Sigma-Aldrich Cat# M1830000 Isoproterenol Cayman Chemical Cat# 15592 DMSO Sigma-Aldrich Cat# D2650 DAPI Sigma-Aldrich Cat# 09564 Fluo-4 calcium dye ThermoFisher Cat# F14201 Critical commercial assays Precision gRNA Synthesis kit ThermoFisher Cat# A29377 ddPCRTM Supermix Biorad Cat# 1863024 LipofectamineTM Stem Transfection Reagent ThermoFisher Cat# STEM00015 Myco Alert kit Lonza Cat# LT07-418 ATPase assay kit Abcam Cat# Ab234055 Deposited data Whole genome sequencing European Genome-Phenome Archive (EGA) https://ega-archive.org/; EGAS00001004929 Experimental models: Cell lines PGPC17 Hildebrandt et al.27 PGPC17 MYH7 V606M patient cell line This study 80 MYH7 R453C patient cell line This study 81 MYBPC3 G148R patient cell line This study 82 MYBPC3 P955fs TNNI3 A157V patient cell line This study 83 MYH7 V606M corrected cell line This study 80C MYH7 R453C corrected cell line This study 81C MYBPC3 G148R corrected cell line This study 82C MYBPC3 P955fs corrected cell line This study 83C MYBPC3 G148R edited line This study G148R MYBPC3 R502W edited line This study R502W MYBPC3 Comp. het. edited line This study Comp. het.
Article Title: Myosin inhibitor reverses hypertrophic cardiomyopathy in genotypically diverse pediatric iPSC-cardiomyocytes to mirror variant correction.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies OCT3/4 antibody PE-conjugated BD Biosciences Cat# 561556; RRID:AB_10714783 Sox2 PerCP-Cy5.5- conjugated BD Biosciences Cat# 561506; RRID:AB_10646039 SSEA-4 V450-conjugated BD Biosciences Cat# 561156; RRID:AB_10896140 TRA-1-60 AF647-conjugated Biolegend Cat# 330605; RRID:AB_1227813 a-actinin antibody Abcam Cat# Ab9465; RRID:AB_307264 MLC2v antibody Abcam Cat# Ab79935; RRID:AB_1952220 Actin Antibody R&D Systems Cat# MAB93081; RRID:AB_3068544 ANP antibody R&D Systems Cat# AF3366; RRID:AB_2915938 TNNI3 antibody R&D Systems Cat# MAB68871; RRID:AB_3068546 MYH7 antibody R&D Systems Cat# MAB9096; RRID:AB_2915972 MYBPC3 antibody Abcam Cat# Ab108522; RRID:AB_10865856 GAPDH antibody Abcam Cat# Ab8245; RRID:AB_2107448 Anti-rabbit 555 secondary antibody ThermoFisher Cat# A-21429; RRID:AB_2535850 Anti-mouse 488 secondary antibody ThermoFisher Cat# A-21121; RRID:AB_2535764 Anti-rabbit 647 secondary antibody ThermoFisher Cat# A-32795; RRID:AB_2762835 Anti-mouse 647 secondary antibody ThermoFisher Cat# A-21240; RRID:AB_2535809 Anti-mouse IgG HRP conjugated R&D Systems Cat# HAF018; RRID: AB_573130 Anti-rabbit IgG HRP conjugated R&D Systems Cat# HAF008; RRID:AB_357235 Biological samples Patient fibroblasts or lymphoblasts Sickkids Heart Center Biobank https://theheartcentrebiobank.com/ Chemicals, peptides, and recombinant proteins Puromycin Sigma-Aldrich Cat# P8833 Fibronectin ThermoFisher Cat# 33016015 Gelatin Sigma-Aldrich Cat# G9391 Matrigel Corning Cat# 354230 Collagen Corning Cat# 354249 PSC Cardiomyocyte Differentiation Kit ThermoFisher Cat# A2921201 STEMdiff cardiomyocyte differentiation kit STEMCELL Technologies Cat# 05010 STEMdiff cardiomyocyte dissociation kit STEMCELL Technologies Cat# 05025 iCell cardiomyocyte maintenance medium FUJIFILM Cellular Dymamics Cat# M1003 RPMI ThermoFisher Cat# 11875-093 B-27 ThermoFisher Cat# 17504-044 StemPro-34 ThermoFisher Cat# 10639011 CHIR-99021 MedChemExpress Cat# HY-10182 cOmplete protease inhibitor cocktail Sigma-Aldrich Cat# 11836170001 Protein A-agarose beads Cell Signaling Technology Cat# 9863 Protein G agarose beads Cell Signaling Technology Cat# 37478 Pierce ECL Western Blotting Substrate ThermoFisher Cat# 32109 (Continued on next page) Cell Reports Medicine 5, 101520, May 21, 2024 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER MYK-461 MedChemExpress Cat# SAR439152 Aficamten Cayman Chemical Cat# 36304 Verapamil Sigma-Aldrich Cat# V45629 Metoprolol Sigma-Aldrich Cat# M1830000 Isoproterenol Cayman Chemical Cat# 15592 DMSO Sigma-Aldrich Cat# D2650 DAPI Sigma-Aldrich Cat# 09564 Fluo-4 calcium dye ThermoFisher Cat# F14201 Critical commercial assays Precision gRNA Synthesis kit ThermoFisher Cat# A29377 ddPCRTM Supermix Biorad Cat# 1863024 LipofectamineTM Stem Transfection Reagent ThermoFisher Cat# STEM00015 Myco Alert kit Lonza Cat# LT07-418 ATPase assay kit Abcam Cat# Ab234055 Deposited data Whole genome sequencing European Genome-Phenome Archive (EGA) https://ega-archive.org/; EGAS00001004929 Experimental models: Cell lines PGPC17 Hildebrandt et al.27 PGPC17 MYH7 V606M patient cell line This study 80 MYH7 R453C patient cell line This study 81 MYBPC3 G148R patient cell line This study 82 MYBPC3 P955fs TNNI3 A157V patient cell line This study 83 MYH7 V606M corrected cell line This study 80C MYH7 R453C corrected cell line This study 81C MYBPC3 G148R corrected cell line This study 82C MYBPC3 P955fs corrected cell line This study 83C MYBPC3 G148R edited line This study G148R MYBPC3 R502W edited line This study R502W MYBPC3 Comp. het. edited line This study Comp. het.
Protease Inhibitor:Article Title: Myosin inhibitor reverses hypertrophic cardiomyopathy in genotypically diverse pediatric iPSC-cardiomyocytes to mirror variant correction
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies OCT3/4 antibody PE-conjugated BD Biosciences Cat# 561556; RRID:AB_10714783 Sox2 PerCP-Cy5.5- conjugated BD Biosciences Cat# 561506; RRID:AB_10646039 SSEA-4 V450-conjugated BD Biosciences Cat# 561156; RRID:AB_10896140 TRA-1-60 AF647-conjugated Biolegend Cat# 330605; RRID:AB_1227813 a-actinin antibody Abcam Cat# Ab9465; RRID:AB_307264 MLC2v antibody Abcam Cat# Ab79935; RRID:AB_1952220 Actin Antibody R&D Systems Cat# MAB93081; RRID:AB_3068544 ANP antibody R&D Systems Cat# AF3366; RRID:AB_2915938 TNNI3 antibody R&D Systems Cat# MAB68871; RRID:AB_3068546 MYH7 antibody R&D Systems Cat# MAB9096; RRID:AB_2915972 MYBPC3 antibody Abcam Cat# Ab108522; RRID:AB_10865856 GAPDH antibody Abcam Cat# Ab8245; RRID:AB_2107448 Anti-rabbit 555 secondary antibody ThermoFisher Cat# A-21429; RRID:AB_2535850 Anti-mouse 488 secondary antibody ThermoFisher Cat# A-21121; RRID:AB_2535764 Anti-rabbit 647 secondary antibody ThermoFisher Cat# A-32795; RRID:AB_2762835 Anti-mouse 647 secondary antibody ThermoFisher Cat# A-21240; RRID:AB_2535809 Anti-mouse IgG HRP conjugated R&D Systems Cat# HAF018; RRID: AB_573130 Anti-rabbit IgG HRP conjugated R&D Systems Cat# HAF008; RRID:AB_357235 Biological samples Patient fibroblasts or lymphoblasts Sickkids Heart Center Biobank https://theheartcentrebiobank.com/ Chemicals, peptides, and recombinant proteins Puromycin Sigma-Aldrich Cat# P8833 Fibronectin ThermoFisher Cat# 33016015 Gelatin Sigma-Aldrich Cat# G9391 Matrigel Corning Cat# 354230 Collagen Corning Cat# 354249 PSC Cardiomyocyte Differentiation Kit ThermoFisher Cat# A2921201 STEMdiff cardiomyocyte differentiation kit STEMCELL Technologies Cat# 05010 STEMdiff cardiomyocyte dissociation kit STEMCELL Technologies Cat# 05025 iCell cardiomyocyte maintenance medium FUJIFILM Cellular Dymamics Cat# M1003 RPMI ThermoFisher Cat# 11875-093 B-27 ThermoFisher Cat# 17504-044 StemPro-34 ThermoFisher Cat# 10639011 CHIR-99021 MedChemExpress Cat# HY-10182 cOmplete protease inhibitor cocktail Sigma-Aldrich Cat# 11836170001 Protein A-agarose beads Cell Signaling Technology Cat# 9863 Protein G agarose beads Cell Signaling Technology Cat# 37478 Pierce ECL Western Blotting Substrate ThermoFisher Cat# 32109 (Continued on next page) Cell Reports Medicine 5, 101520, May 21, 2024 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER MYK-461 MedChemExpress Cat# SAR439152 Aficamten Cayman Chemical Cat# 36304 Verapamil Sigma-Aldrich Cat# V45629 Metoprolol Sigma-Aldrich Cat# M1830000 Isoproterenol Cayman Chemical Cat# 15592 DMSO Sigma-Aldrich Cat# D2650 DAPI Sigma-Aldrich Cat# 09564 Fluo-4 calcium dye ThermoFisher Cat# F14201 Critical commercial assays Precision gRNA Synthesis kit ThermoFisher Cat# A29377 ddPCRTM Supermix Biorad Cat# 1863024 LipofectamineTM Stem Transfection Reagent ThermoFisher Cat# STEM00015 Myco Alert kit Lonza Cat# LT07-418 ATPase assay kit Abcam Cat# Ab234055 Deposited data Whole genome sequencing European Genome-Phenome Archive (EGA) https://ega-archive.org/; EGAS00001004929 Experimental models: Cell lines PGPC17 Hildebrandt et al.27 PGPC17 MYH7 V606M patient cell line This study 80 MYH7 R453C patient cell line This study 81 MYBPC3 G148R patient cell line This study 82 MYBPC3 P955fs TNNI3 A157V patient cell line This study 83 MYH7 V606M corrected cell line This study 80C MYH7 R453C corrected cell line This study 81C MYBPC3 G148R corrected cell line This study 82C MYBPC3 P955fs corrected cell line This study 83C MYBPC3 G148R edited line This study G148R MYBPC3 R502W edited line This study R502W MYBPC3 Comp. het. edited line This study Comp. het.
Article Title: Myosin inhibitor reverses hypertrophic cardiomyopathy in genotypically diverse pediatric iPSC-cardiomyocytes to mirror variant correction.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies OCT3/4 antibody PE-conjugated BD Biosciences Cat# 561556; RRID:AB_10714783 Sox2 PerCP-Cy5.5- conjugated BD Biosciences Cat# 561506; RRID:AB_10646039 SSEA-4 V450-conjugated BD Biosciences Cat# 561156; RRID:AB_10896140 TRA-1-60 AF647-conjugated Biolegend Cat# 330605; RRID:AB_1227813 a-actinin antibody Abcam Cat# Ab9465; RRID:AB_307264 MLC2v antibody Abcam Cat# Ab79935; RRID:AB_1952220 Actin Antibody R&D Systems Cat# MAB93081; RRID:AB_3068544 ANP antibody R&D Systems Cat# AF3366; RRID:AB_2915938 TNNI3 antibody R&D Systems Cat# MAB68871; RRID:AB_3068546 MYH7 antibody R&D Systems Cat# MAB9096; RRID:AB_2915972 MYBPC3 antibody Abcam Cat# Ab108522; RRID:AB_10865856 GAPDH antibody Abcam Cat# Ab8245; RRID:AB_2107448 Anti-rabbit 555 secondary antibody ThermoFisher Cat# A-21429; RRID:AB_2535850 Anti-mouse 488 secondary antibody ThermoFisher Cat# A-21121; RRID:AB_2535764 Anti-rabbit 647 secondary antibody ThermoFisher Cat# A-32795; RRID:AB_2762835 Anti-mouse 647 secondary antibody ThermoFisher Cat# A-21240; RRID:AB_2535809 Anti-mouse IgG HRP conjugated R&D Systems Cat# HAF018; RRID: AB_573130 Anti-rabbit IgG HRP conjugated R&D Systems Cat# HAF008; RRID:AB_357235 Biological samples Patient fibroblasts or lymphoblasts Sickkids Heart Center Biobank https://theheartcentrebiobank.com/ Chemicals, peptides, and recombinant proteins Puromycin Sigma-Aldrich Cat# P8833 Fibronectin ThermoFisher Cat# 33016015 Gelatin Sigma-Aldrich Cat# G9391 Matrigel Corning Cat# 354230 Collagen Corning Cat# 354249 PSC Cardiomyocyte Differentiation Kit ThermoFisher Cat# A2921201 STEMdiff cardiomyocyte differentiation kit STEMCELL Technologies Cat# 05010 STEMdiff cardiomyocyte dissociation kit STEMCELL Technologies Cat# 05025 iCell cardiomyocyte maintenance medium FUJIFILM Cellular Dymamics Cat# M1003 RPMI ThermoFisher Cat# 11875-093 B-27 ThermoFisher Cat# 17504-044 StemPro-34 ThermoFisher Cat# 10639011 CHIR-99021 MedChemExpress Cat# HY-10182 cOmplete protease inhibitor cocktail Sigma-Aldrich Cat# 11836170001 Protein A-agarose beads Cell Signaling Technology Cat# 9863 Protein G agarose beads Cell Signaling Technology Cat# 37478 Pierce ECL Western Blotting Substrate ThermoFisher Cat# 32109 (Continued on next page) Cell Reports Medicine 5, 101520, May 21, 2024 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER MYK-461 MedChemExpress Cat# SAR439152 Aficamten Cayman Chemical Cat# 36304 Verapamil Sigma-Aldrich Cat# V45629 Metoprolol Sigma-Aldrich Cat# M1830000 Isoproterenol Cayman Chemical Cat# 15592 DMSO Sigma-Aldrich Cat# D2650 DAPI Sigma-Aldrich Cat# 09564 Fluo-4 calcium dye ThermoFisher Cat# F14201 Critical commercial assays Precision gRNA Synthesis kit ThermoFisher Cat# A29377 ddPCRTM Supermix Biorad Cat# 1863024 LipofectamineTM Stem Transfection Reagent ThermoFisher Cat# STEM00015 Myco Alert kit Lonza Cat# LT07-418 ATPase assay kit Abcam Cat# Ab234055 Deposited data Whole genome sequencing European Genome-Phenome Archive (EGA) https://ega-archive.org/; EGAS00001004929 Experimental models: Cell lines PGPC17 Hildebrandt et al.27 PGPC17 MYH7 V606M patient cell line This study 80 MYH7 R453C patient cell line This study 81 MYBPC3 G148R patient cell line This study 82 MYBPC3 P955fs TNNI3 A157V patient cell line This study 83 MYH7 V606M corrected cell line This study 80C MYH7 R453C corrected cell line This study 81C MYBPC3 G148R corrected cell line This study 82C MYBPC3 P955fs corrected cell line This study 83C MYBPC3 G148R edited line This study G148R MYBPC3 R502W edited line This study R502W MYBPC3 Comp. het. edited line This study Comp. het.
Western Blot:Article Title: Myosin inhibitor reverses hypertrophic cardiomyopathy in genotypically diverse pediatric iPSC-cardiomyocytes to mirror variant correction
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies OCT3/4 antibody PE-conjugated BD Biosciences Cat# 561556; RRID:AB_10714783 Sox2 PerCP-Cy5.5- conjugated BD Biosciences Cat# 561506; RRID:AB_10646039 SSEA-4 V450-conjugated BD Biosciences Cat# 561156; RRID:AB_10896140 TRA-1-60 AF647-conjugated Biolegend Cat# 330605; RRID:AB_1227813 a-actinin antibody Abcam Cat# Ab9465; RRID:AB_307264 MLC2v antibody Abcam Cat# Ab79935; RRID:AB_1952220 Actin Antibody R&D Systems Cat# MAB93081; RRID:AB_3068544 ANP antibody R&D Systems Cat# AF3366; RRID:AB_2915938 TNNI3 antibody R&D Systems Cat# MAB68871; RRID:AB_3068546 MYH7 antibody R&D Systems Cat# MAB9096; RRID:AB_2915972 MYBPC3 antibody Abcam Cat# Ab108522; RRID:AB_10865856 GAPDH antibody Abcam Cat# Ab8245; RRID:AB_2107448 Anti-rabbit 555 secondary antibody ThermoFisher Cat# A-21429; RRID:AB_2535850 Anti-mouse 488 secondary antibody ThermoFisher Cat# A-21121; RRID:AB_2535764 Anti-rabbit 647 secondary antibody ThermoFisher Cat# A-32795; RRID:AB_2762835 Anti-mouse 647 secondary antibody ThermoFisher Cat# A-21240; RRID:AB_2535809 Anti-mouse IgG HRP conjugated R&D Systems Cat# HAF018; RRID: AB_573130 Anti-rabbit IgG HRP conjugated R&D Systems Cat# HAF008; RRID:AB_357235 Biological samples Patient fibroblasts or lymphoblasts Sickkids Heart Center Biobank https://theheartcentrebiobank.com/ Chemicals, peptides, and recombinant proteins Puromycin Sigma-Aldrich Cat# P8833 Fibronectin ThermoFisher Cat# 33016015 Gelatin Sigma-Aldrich Cat# G9391 Matrigel Corning Cat# 354230 Collagen Corning Cat# 354249 PSC Cardiomyocyte Differentiation Kit ThermoFisher Cat# A2921201 STEMdiff cardiomyocyte differentiation kit STEMCELL Technologies Cat# 05010 STEMdiff cardiomyocyte dissociation kit STEMCELL Technologies Cat# 05025 iCell cardiomyocyte maintenance medium FUJIFILM Cellular Dymamics Cat# M1003 RPMI ThermoFisher Cat# 11875-093 B-27 ThermoFisher Cat# 17504-044 StemPro-34 ThermoFisher Cat# 10639011 CHIR-99021 MedChemExpress Cat# HY-10182 cOmplete protease inhibitor cocktail Sigma-Aldrich Cat# 11836170001 Protein A-agarose beads Cell Signaling Technology Cat# 9863 Protein G agarose beads Cell Signaling Technology Cat# 37478 Pierce ECL Western Blotting Substrate ThermoFisher Cat# 32109 (Continued on next page) Cell Reports Medicine 5, 101520, May 21, 2024 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER MYK-461 MedChemExpress Cat# SAR439152 Aficamten Cayman Chemical Cat# 36304 Verapamil Sigma-Aldrich Cat# V45629 Metoprolol Sigma-Aldrich Cat# M1830000 Isoproterenol Cayman Chemical Cat# 15592 DMSO Sigma-Aldrich Cat# D2650 DAPI Sigma-Aldrich Cat# 09564 Fluo-4 calcium dye ThermoFisher Cat# F14201 Critical commercial assays Precision gRNA Synthesis kit ThermoFisher Cat# A29377 ddPCRTM Supermix Biorad Cat# 1863024 LipofectamineTM Stem Transfection Reagent ThermoFisher Cat# STEM00015 Myco Alert kit Lonza Cat# LT07-418 ATPase assay kit Abcam Cat# Ab234055 Deposited data Whole genome sequencing European Genome-Phenome Archive (EGA) https://ega-archive.org/; EGAS00001004929 Experimental models: Cell lines PGPC17 Hildebrandt et al.27 PGPC17 MYH7 V606M patient cell line This study 80 MYH7 R453C patient cell line This study 81 MYBPC3 G148R patient cell line This study 82 MYBPC3 P955fs TNNI3 A157V patient cell line This study 83 MYH7 V606M corrected cell line This study 80C MYH7 R453C corrected cell line This study 81C MYBPC3 G148R corrected cell line This study 82C MYBPC3 P955fs corrected cell line This study 83C MYBPC3 G148R edited line This study G148R MYBPC3 R502W edited line This study R502W MYBPC3 Comp. het. edited line This study Comp. het.
Article Title: Myosin inhibitor reverses hypertrophic cardiomyopathy in genotypically diverse pediatric iPSC-cardiomyocytes to mirror variant correction.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies OCT3/4 antibody PE-conjugated BD Biosciences Cat# 561556; RRID:AB_10714783 Sox2 PerCP-Cy5.5- conjugated BD Biosciences Cat# 561506; RRID:AB_10646039 SSEA-4 V450-conjugated BD Biosciences Cat# 561156; RRID:AB_10896140 TRA-1-60 AF647-conjugated Biolegend Cat# 330605; RRID:AB_1227813 a-actinin antibody Abcam Cat# Ab9465; RRID:AB_307264 MLC2v antibody Abcam Cat# Ab79935; RRID:AB_1952220 Actin Antibody R&D Systems Cat# MAB93081; RRID:AB_3068544 ANP antibody R&D Systems Cat# AF3366; RRID:AB_2915938 TNNI3 antibody R&D Systems Cat# MAB68871; RRID:AB_3068546 MYH7 antibody R&D Systems Cat# MAB9096; RRID:AB_2915972 MYBPC3 antibody Abcam Cat# Ab108522; RRID:AB_10865856 GAPDH antibody Abcam Cat# Ab8245; RRID:AB_2107448 Anti-rabbit 555 secondary antibody ThermoFisher Cat# A-21429; RRID:AB_2535850 Anti-mouse 488 secondary antibody ThermoFisher Cat# A-21121; RRID:AB_2535764 Anti-rabbit 647 secondary antibody ThermoFisher Cat# A-32795; RRID:AB_2762835 Anti-mouse 647 secondary antibody ThermoFisher Cat# A-21240; RRID:AB_2535809 Anti-mouse IgG HRP conjugated R&D Systems Cat# HAF018; RRID: AB_573130 Anti-rabbit IgG HRP conjugated R&D Systems Cat# HAF008; RRID:AB_357235 Biological samples Patient fibroblasts or lymphoblasts Sickkids Heart Center Biobank https://theheartcentrebiobank.com/ Chemicals, peptides, and recombinant proteins Puromycin Sigma-Aldrich Cat# P8833 Fibronectin ThermoFisher Cat# 33016015 Gelatin Sigma-Aldrich Cat# G9391 Matrigel Corning Cat# 354230 Collagen Corning Cat# 354249 PSC Cardiomyocyte Differentiation Kit ThermoFisher Cat# A2921201 STEMdiff cardiomyocyte differentiation kit STEMCELL Technologies Cat# 05010 STEMdiff cardiomyocyte dissociation kit STEMCELL Technologies Cat# 05025 iCell cardiomyocyte maintenance medium FUJIFILM Cellular Dymamics Cat# M1003 RPMI ThermoFisher Cat# 11875-093 B-27 ThermoFisher Cat# 17504-044 StemPro-34 ThermoFisher Cat# 10639011 CHIR-99021 MedChemExpress Cat# HY-10182 cOmplete protease inhibitor cocktail Sigma-Aldrich Cat# 11836170001 Protein A-agarose beads Cell Signaling Technology Cat# 9863 Protein G agarose beads Cell Signaling Technology Cat# 37478 Pierce ECL Western Blotting Substrate ThermoFisher Cat# 32109 (Continued on next page) Cell Reports Medicine 5, 101520, May 21, 2024 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER MYK-461 MedChemExpress Cat# SAR439152 Aficamten Cayman Chemical Cat# 36304 Verapamil Sigma-Aldrich Cat# V45629 Metoprolol Sigma-Aldrich Cat# M1830000 Isoproterenol Cayman Chemical Cat# 15592 DMSO Sigma-Aldrich Cat# D2650 DAPI Sigma-Aldrich Cat# 09564 Fluo-4 calcium dye ThermoFisher Cat# F14201 Critical commercial assays Precision gRNA Synthesis kit ThermoFisher Cat# A29377 ddPCRTM Supermix Biorad Cat# 1863024 LipofectamineTM Stem Transfection Reagent ThermoFisher Cat# STEM00015 Myco Alert kit Lonza Cat# LT07-418 ATPase assay kit Abcam Cat# Ab234055 Deposited data Whole genome sequencing European Genome-Phenome Archive (EGA) https://ega-archive.org/; EGAS00001004929 Experimental models: Cell lines PGPC17 Hildebrandt et al.27 PGPC17 MYH7 V606M patient cell line This study 80 MYH7 R453C patient cell line This study 81 MYBPC3 G148R patient cell line This study 82 MYBPC3 P955fs TNNI3 A157V patient cell line This study 83 MYH7 V606M corrected cell line This study 80C MYH7 R453C corrected cell line This study 81C MYBPC3 G148R corrected cell line This study 82C MYBPC3 P955fs corrected cell line This study 83C MYBPC3 G148R edited line This study G148R MYBPC3 R502W edited line This study R502W MYBPC3 Comp. het. edited line This study Comp. het.
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