Fig. 7. Hip1r localizes to the plasma membrane and cytoplasm and displays an interaction with epsin. Growing cells were fixed and stained with antibody against Hip1r. (A) Cells were viewed by DIC microscopy. (B) Cells stained with antibody against Hip1r show bright Hip1r puncta at the cell cortex when viewed by fluorescence microscopy. (C-E) Focusing on the surface of cells expressing a clathrin-GFP construct (clathrin:GFP) shows partial colocalization of clathrin (green) with Hip1r (anti-Hip1r, red). Examples are indicated by arrowheads (E). (F-H) Cells expressing an epsin-GFP fusion protein (epsin:GFP, green) and stained with antibody against Hip1r (anti-Hip1r, red) exhibit extensive colocalization at the plasma membrane (arrowheads). Occasionally, puncta contain epsin but not Hip1r (arrow). Bar, 10 µm.