Movie 2.

ERMES foci form de novo. Cells expressing Mdm12-Halo, the mitochondrial matrix marker MitoRed, and the ER marker GFP-HDEL, were grown to mid-log phase, labeled with JFX650, adhered to a ConA treated confocal dish, and imaged. The movie depicts an event where a new Mdm12 focus forms. The movie is a max projection of full Z-stacks to demonstrate that no Mdm12 focus was present in the cell at that location prior to formation. A merged image of all fluorescence channels is shown on top and the Mdm12-Halo channel is shown in grayscale below. Images from this movie are shown in Fig. 3A. Scale bar 2 μm. Time is in min:s.