Movie 2

3D reconstruction showing the Z-stack images with step size = 0.12 μm. Z-stack shows the bottom plane and the upper plane of the image showing the presence of the TNTs connecting a RAW/LR5 macrophage with MTLn3 tumor cells. Cells were fixed and stained for WGA (green) and F-actin (magenta). homotypic TNTs (indicated by white arrows) are often observed between two macrophages or two tumor cells. Images were acquired using a Nikon Structured Illumination N-SIM system on an inverted Nikon ECLIPSE Ti-E equipped with a 100× 1.49 NA objective. Analysis performed with ImageJ (Rasband, W.S., ImageJ, U. S. National Institutes of Health, Bethesda, Maryland, USA, http://imagej.nih.gov/ij/, 1997–2016).