2.11. Construction of the Ndusf1 siRNA and Overexpression Vectors and Ang II Treatment

The siRNAs targeting Ndusf1 (si-Ndusf1) and the corresponding scrambled siRNAs were designed and synthesized by RiboBio (Guangzhou, China). The vector for Ndusf1 overexpression was constructed by cloning the full-length Ndusf1 sequence into the pcDNA3.1 vector, and the empty pcDNA3.1 vector was used as the corresponding negative control. All plasmids were purchased from RiboBio. For transfections, rat cardiomyocytes were seeded in 12-well plates and cultured for 24 h; then, cardiomyocytes were transfected with various plasmids or siRNAs by using Lipofectamine 2000 reagent (Invitrogen, Carlsbad, USA) according to the manufacturer's protocol. Cardiomyocytes were collected for the experiments 24 h after the transfection. For angiotensin II (Ang II; Sigma-Aldrich) treatment, cardiomyocytes were seeded in 12-well plates and cultured for 24 h; then, cardiomyocytes were treated with 100 nM Ang II for 24 h and harvested for subsequent experiments.