The ACADVL gene encodes for very long-chain acyl-CoA dehydrogenase (VLCAD), which functions within mitochondria and is essential for fatty acid oxidation. HCFs were prepared for western blot analysis of VLCAD, caspase-3 (CASP3), cytochrome c (Cyto C), lamin B1, β-actin, and Arp2 with the internal reference protein glyceraldehyde 3-phosphate dehydrogenase (GAPDH) before knockdown of the ACADVL gene. The above proteins were quantified again after knockdown of ACADVL. Detailed methods of the western blotting and knockdown procedure are provided in Additional file 2 .
Knockdown of DNMT1 leads to generally decreased DNA methylation and activates cascades of genotoxic stress [31 (link)] in cells, resulting in signal transduction unrelated to cardiac fibrosis. Thus, we preferred to downregulate the ACADVL gene expression to simulate the HIIT-associated inhibition of human cardiac fibroblast activities.
Knockdown of DNMT1 leads to generally decreased DNA methylation and activates cascades of genotoxic stress [31 (link)] in cells, resulting in signal transduction unrelated to cardiac fibrosis. Thus, we preferred to downregulate the ACADVL gene expression to simulate the HIIT-associated inhibition of human cardiac fibroblast activities.
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