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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV220410	1/2	Mus musculus	heart tissue	(d)(U)C	Keat Tham, Yow	2023	56%
Study summary Full title Estrogen receptor alpha deficiency in cardiomyocytes reprograms the heart-derived extracellular vesicle proteome and induces obesity in female mice All authors Yow Keat Tham, Bianca C. Bernardo, Bethany Claridge, Gunes S. Yildiz, Liesel Min-Linn Woon, Simon Bond, Haoyun Fang, Jenny Y. Y. Ooi, Aya Matsumoto, Jieting Luo, Celeste M. K. Tai, Claudia A. Harmawan, Helen Kiriazis, Daniel G. Donner, Natalie A. Mellett, E. Dale Abel, Sohaib A. Khan, David P. De Souza, Sheik Nadeem Elahee Doomun, Kevin Liu, Ruidong Xiang, Manika Singh, Michael Inouye, Peter J. Meikle, Kate L. Weeks, Brian G. Drew, David W. Greening & Julie R. McMullen Journal Abstract Dysregulation of estrogen receptor alpha (ERα) has been linked with increased metabolic and cardiov (show more...)Dysregulation of estrogen receptor alpha (ERα) has been linked with increased metabolic and cardiovascular disease risk. Here, we generate and characterize cardiomyocyte-specific ERα knockout (ERαHKO) mice to assess the role of ERα in the heart. The most striking phenotype was obesity in female ERαHKO but not male ERαHKO mice. Female ERαHKO mice showed cardiac dysfunction, mild glucose and insulin intolerance and reduced ERα gene expression in skeletal muscle and white adipose tissue. Transcriptomic, proteomic, lipidomic and metabolomic analyses revealed evidence of contractile and/or metabolic dysregulation in heart, skeletal muscle and white adipose tissue. We show that heart-derived extracellular vesicles from female ERαHKO mice contain a distinct proteome associated with lipid and metabolic regulation, and have the capacity to metabolically reprogram the target skeletal myocyte proteome with functional impacts on glycolytic capacity and reserve. This multi-omics study uncovers a cardiac-initiated and sex-specific cardiometabolic phenotype regulated by ERα and provides insights into extracellular vesicle-mediated interorgan communication. (hide) EV-METRIC 56% (57th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type heart tissue Sample origin Control condition Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Protein markers EV: Alix/ CD9/ CD63/ CD81/ TSG101 non-EV: Calreticulin/ Albumin/ fga/ hba Proteomics yes Show all info Study aim Function/Identification of content (omics approaches) Sample Species Mus musculus Sample Type heart tissue Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: rotor type TLA-100 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 1 Wash: time (min) 60 Wash: Rotor Type TLA-100 Wash: speed (g) 100000 Characterization: Protein analysis Protein Concentration Method microBCA Protein Yield (µg) total yield (from tissue starting sample, intact heart) Western Blot Detected EV-associated proteins Alix/ CD9/ CD63/ CD81/ TSG101 Not detected contaminants Calreticulin Proteomics database PRIDE proteomics database Not detected contaminants Albumin/ fga/ hba Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) 185 EV concentration Yes Particle yield particles per milliliter of starting sample: 9.00E+08

	EV220410	2/2	Mus musculus	heart tissue	(d)(U)C	Keat Tham, Yow	2023	56%
Study summary Full title Estrogen receptor alpha deficiency in cardiomyocytes reprograms the heart-derived extracellular vesicle proteome and induces obesity in female mice All authors Yow Keat Tham, Bianca C. Bernardo, Bethany Claridge, Gunes S. Yildiz, Liesel Min-Linn Woon, Simon Bond, Haoyun Fang, Jenny Y. Y. Ooi, Aya Matsumoto, Jieting Luo, Celeste M. K. Tai, Claudia A. Harmawan, Helen Kiriazis, Daniel G. Donner, Natalie A. Mellett, E. Dale Abel, Sohaib A. Khan, David P. De Souza, Sheik Nadeem Elahee Doomun, Kevin Liu, Ruidong Xiang, Manika Singh, Michael Inouye, Peter J. Meikle, Kate L. Weeks, Brian G. Drew, David W. Greening & Julie R. McMullen Journal Abstract Dysregulation of estrogen receptor alpha (ERα) has been linked with increased metabolic and cardiov (show more...)Dysregulation of estrogen receptor alpha (ERα) has been linked with increased metabolic and cardiovascular disease risk. Here, we generate and characterize cardiomyocyte-specific ERα knockout (ERαHKO) mice to assess the role of ERα in the heart. The most striking phenotype was obesity in female ERαHKO but not male ERαHKO mice. Female ERαHKO mice showed cardiac dysfunction, mild glucose and insulin intolerance and reduced ERα gene expression in skeletal muscle and white adipose tissue. Transcriptomic, proteomic, lipidomic and metabolomic analyses revealed evidence of contractile and/or metabolic dysregulation in heart, skeletal muscle and white adipose tissue. We show that heart-derived extracellular vesicles from female ERαHKO mice contain a distinct proteome associated with lipid and metabolic regulation, and have the capacity to metabolically reprogram the target skeletal myocyte proteome with functional impacts on glycolytic capacity and reserve. This multi-omics study uncovers a cardiac-initiated and sex-specific cardiometabolic phenotype regulated by ERα and provides insights into extracellular vesicle-mediated interorgan communication. (hide) EV-METRIC 56% (57th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type heart tissue Sample origin ERa KO Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Protein markers EV: Alix/ CD63/ CD81/ TSG101 non-EV: Calreticulin/ fga/ hba Proteomics yes Show all info Study aim Function/Identification of content (omics approaches) Sample Species Mus musculus Sample Type heart tissue Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: rotor type TLA-100 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) 1 Wash: time (min) 60 Wash: Rotor Type TLA-100 Wash: speed (g) 100000 Characterization: Protein analysis Protein Concentration Method microBCA Protein Yield (µg) total yield (from tissue starting sample, intact heart) Western Blot Detected EV-associated proteins Alix/ CD63/ CD81/ TSG101 Not detected contaminants Calreticulin Proteomics database PRIDE proteomics database Not detected contaminants fga/ hba Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) 185 EV concentration Yes Particle yield particles per milliliter of starting sample: 9.00E+08

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EV-TRACK ID	EV220410
species	Mus musculus
sample type	heart tissue
condition	Control condition	ERa KO
separation protocol	dUC	dUC
Exp. nr.	1	2
EV-METRIC %	56	56