Dynamin2 and Endophilin A2 Interactions

Genome-edited SK-MEL-2 cells expressing the endogenous dynamin2 with a GFP tag (kind gift from David Drubin lab, UC Berkeley, USA; [Doyon et al., 2011 (link)]) were transfected using 0.5 µg of plasmids containing human endophilin A2 with C-terminal TagRFP-T under the control of the CMV promoter with X-tremeGENE HP DNA Transfection Reagent (06366236001, Roche Diagnostics GmbH, Mannheim, Germany) according to the manufacturer’s protocol. The endophilin plasmid was kindly provided by Emmanuel Boucrot, University College London, UK. For the recovery experiments, human dynamin2 cherry (kindly provided by Christien Merrifield, LEBS, Gif-Sur-Yvette, France) was overexpressed together with a GFP version of human endophilinA2 (kindly provided by Emmanuel Boucrot [Boucrot et al., 2015 (link)]). Images of the cells were acquired on a microscope (Nikon Eclipse T1, Nikon Tokyo, Japan) with a 100x oil objective (Nikon CFI Apo TIRF 100x NA 1.4) using total internal reflection fluorescence (TIRF).