Three-dimensional distribution of PTEN (green) in relation to actin waves (red). Wave-forming cells expressing GFP-PTEN and mRFP-LimEΔ were scanned by spinning-disc confocal microscopy. A, three optical sections from a z-stack scanned at the substrate (left), and 600 nm (middle), or 1200 nm ahead of the substrate (right). This series indicates that PTEN in the external area of the substrate-attached surface is contiguous with PTEN bound to the membrane of the non-attached surface (arrowhead). B, three-dimensional PTEN pattern in relation to an actin wave. The reconstruction shows the continuity of the PTEN-rich area at the border of the free and substrate-attached cell surface (arrowhead in the middle panel) and membrane location of the PTEN (bottom panel). To view the inside of the cell, its top has been optically cut off in the bottom panel, and the plane of the cut-off section shown in gray. C, 3D-reconstruction similar to (B). The PTEN-rich border between free and substrate-attached cell surface is seen (arrowheads). A cross-section through the stacks of PTEN images (white frame in the bottom panel) shows the corner of the PTEN-rich area in a side view. Bars, 10 μ m.
Gerisch et al. BMC Cell Biology 2011 12:42 doi:10.1186/1471-2121-12-42