Modulating Mitochondrial Dynamics in HeLa Cells

HeLa cells were treated with 15 µM APE1 inhibitor III (APE1inhIII) (Merck Millipore, Burlingtion, MA, USA) or the solvent DMSO (Merck Millipore, Burlingtion, MA, USA) for 5 h at 37 °C [28 (link)] before mitochondrial replication imaging protocol (mREP) or total DNA isolation was performed. For scavenging of ROS, HeLa cells were treated for 24 h with either 100 µM mitoTempo (Sigma-Aldrich/Merck, St. Louis, MO, USA) [37 (link)] or N-acetyl-L-cysteine (NAC) (Sigma-Aldrich/Merck, St. Louis, MO, USA) [29 (link),38 (link),39 (link),40 (link)] or water as solvent. To block mitochondrial import of proteins, HeLa cells were treated for 48 h with 10 µM mitoBlock-6 (Focus Biomolecules, Plymouth Meeting, PA, USA). To slow down mitochondrial DNA replication, the HeLa cells were treated for 24 h with 10 µM 2′,3′-Dideoxycytidine (ddC) (Sigma-Aldrich/Merck, St. Louis, MO, USA) [41 (link)].