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INFORMACIÓN GENERAL
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1. Título del dataset: [Lax´Lab]_Study: AEOL-Induced NRF2 Activation and DWORF Overexpression Mitigate Myocardial I/R Injury. Quantification of Western blot data – Experimental methodology
2. Autoría:

[Rellenar la información de todos los autores siguiendo el siguiente formato. Repetir el esquema, uno para cada autor.]

	Nombre:Lax Pérez, Antonio Manuel
	Institución:Universidad de Murcia
	Correo electrónico: alax@um.es
	ORCID:


	Nombre:Asensio-López, María del Carmen
	Institución: BioCardio S.L.
	Correo electrónico: mal24027@um.es
	ORCID:
	

3. Fecha de recogida de los datos (fecha única o rango de fechas): 
From 01-02-2023 to 31-03-2024

4. Fecha de depósito de los datos:
21-02-2025

5. Idioma del conjunto de datos:
English

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INFORMACIÓN METODOLÓGICA 
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1.Descripción de la metodología utilizada para generar el conjunto de datos.

Western blotting was used to generate the dataset. Proteins (35 μg) were denatured, separated by SDS-PAGE, and transferred onto a polyvinylidene difluoride (PVDF) membrane (Merck Millipore, USA). To prevent non-specific binding, membranes were blocked with 5% (w/v) BSA in TBST (137 mM NaCl, 20 mM Tris, and 0.1% (v/v) Tween-20, pH 7.6) and incubated overnight at 4ºC with primary antibodies. After extensive washing with TBST, membranes were incubated for 1 hour at room temperature (RT) with the appropriate secondary antibody, followed by enhanced chemiluminescence detection using the Amersham ECL™ Prime Western Blotting Detection Reagent (GE Healthcare, RPN2232). Imaging and quantification of immunoreactive bands were performed using the ChemiDoc XRS+ system and Image Lab software (Bio-Rad Laboratories). Band density was analyzed with Image Lab, and molecular weight was determined by comparing with prestained protein markers (Precision Plus Protein™ Dual Color Standards, Bio-Rad 1610374).

Protein bands were identified based on their predicted molecular weights, supported by prior data and antibody specifications. Antibody specificity was validated through preliminary testing, ensuring minimal off-target binding. The dataset was generated from n=7 mice per group for in vivo assays and n=5 independent experiments per group for in vitro procedures. Two technical replicates were performed per sample to ensure consistency. Each group included several independent Western blots, each derived from distinct biological replicates. Equal protein loading was verified using GAPDH (for cytosolic fractions) and H3 (for nuclear fractions). 


2. Software o instrumentos necesarios para interpretar los datos: 
[Incluir la versión del software. Si hace falta un software específico de acceso restringido, explicar cómo obtenerlo. Valorar si es posible cambiar el conjunto de datos a un formato abierto (recomendado).] 
(*) Densitometry data were analyzed using Image Lab software from Bio-Rad Laboratories (version 6.1), which was used for quantifying Western blot band intensities. Additionally, histogram graphs were generated, and representative Western blot images were included for each experimental condition.

3. Procedimientos seguidos para asegurar la calidad de los datos: Cuestionario validado, revisión por pares.
(*) Loading control: Antibodies against GAPDH (for cytosolic fractions) and H3 (for nuclear fractions) were used to ensure accurate quantification.
Technical and biological replicates: Three technical replicates were performed per sample, with five independent biological replicates per group.
Antibody validation: Antibodies with high specificity and minimal cross-reactivity were selected based on preliminary testing.
Data documentation: Histogram graphs and representative Western blot images were included for each condition.
Peer review: Data were evaluated by independent researchers to ensure reliability.

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ESCTRUCTURA DE LOS ARCHIVOS
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1. Nombres de archivos 
[Mencionar todos los archivos incluidos en el conjunto de datos, con el nombre y la extensión (.csv, .pdf, etc.) de cada fichero].

(*) AEOL increases nuclear translocation of NRF2 and the transcriptional induction of downstream .xlsx
(*) AEOL treatment does not affect PLN phosphorylation.xlsx
(*) AEOL treatment protects hiPSCMs against simulated I_R injury.xlsx  
(*) AEOL-induced DWORF upregulation displaces PLN from SERCA2a.xlsx


2. Formato de los archivos: xlsx


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MÁS INFORMACIÓN
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 [Incluir cualquier otra información sobre el conjunto de datos que no haya quedado reflejada en esta plantilla y que se considere relevante.]