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ZVV was supported by the National Program of Excellence (TAMOP 4.2.4.A/1-11-1-2012-0001) and by the Rosztoczy Foundation. PF was a Szentagothai Fellow of the National Program of Excellence (TAMOP 4.2.4.A/2-11-1-2012-0001). ZG held a \"Janos Bolyai Fellowship\" from the Hungarian Academy of Sciences (awarded in 2013 and 2017). TB is supported by the UNKP-16-3 New National Excellence Program of The Ministry of Human Capacities. PP is supported by the Intramural Research Program of NIAAA. 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Therefore, we aimed to characterize NOX4 expression in human samples from healthy and failing hearts. Explanted human heart samples (left and right ventricular, and septal regions) were obtained from patients suffering from heart failure of ischemic or dilated origin. Control samples were obtained from donor hearts that were not used for transplantation. Deep RNA sequencing of the cardiac transcriptome indicated extensive alternative splicing of the NOX4 gene in heart failure as compared to samples from healthy donor hearts. Long distance PCR analysis with a universal 5'-3' end primer pair, allowing amplification of different splice variants, confirmed the presence of the splice variants. To assess translation of the alternatively spliced transcripts we determined protein expression of NOX4 by using a specific antibody recognizing a conserved region in all variants. Western blot analysis showed up-regulation of the full-length NOX4 in ischemic cardiomyopathy samples and confirmed presence of shorter isoforms both in control and failing samples with disease-associated expression pattern. We describe here for the first time that NOX4 undergoes extensive alternative splicing in human hearts which gives rise to the expression of different enzyme isoforms. 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