Fig. 2. SNX9-induced tubules are elongated from the plasma membrane. (A-C) COS-7 cells expressing GFP-tagged SNX9 was stained with 2 µM CM-DiI (A), 2 µM cholera toxin B Alexa Fluor 594 (B) or 15 µM FM5-95 (C) for 10 minutes on ice and fixed for observation. SNX9-induced tubules (green) and membrane marker stained plasma membranes (red) were mostly colocalized at cell peripheries. High-magnification views are of the regions enclosed by rectangles. Scale bars: 30 µm (low magnification) and 3 µm (insets). (D) Tubules grown from the bottom of the cell in contact with the coverslip were imaged by TIRF microscopy. Arrowheads indicate tubules elongated from SNX9-positive spots at the plasma membrane. Insets are high-magnification views of the regions enclosed in rectangles. Scale bars: 30 µm (low magnification) and 8 µm (insets).