Drp1 Knockdown in HeLa and N27 Cells

Drp1 knockdown in HeLa and N27 cells were performed as previously described [13 (link)]. Briefly, pre-designed siRNA against human DNM1L (gene encoding Drp1) and rat Dnm1l were purchased from Dharmacon Research, Inc (now available through Horizon Discovery). SMARTpool: siGENOME Human DNM1L siRNA (Cat# 10,059) was used for HeLa cells, while SMARTpool: siGENOME Rat Dnm1l siRNA (Cat# 114,114) was used for N27 cells. Each of this product is a mixture of four siRNA targeting a single gene to enhance efficiency and specificity of Drp1-knockdown. Similarly, the scramble control, siGENOME Non-Targeting siRNA Control Pools (Cat# D-001206, Dharmacon Inc.), was designed to have a minimum of four mismatches to all human, mouse and rat genes, and confirmed by the manufacturer to have minimal targeting by genome-wide microarray analysis. To monitor LC3 levels in N27 cells, mCherry-hLC3B-pcDNA3.1 plasmid was used. All the transfection was performed using Lipofectamine™ 3000 (Thermo Fisher Scientific) according to the manufacturer’s instructions.