Fig. 2. INPP5B does not colocalise with clathrin or bind to clathrin or AP-2. (A) HeLa cells transiently expressing GFP-INPP5B (green) were labelled with antibodies against the clathrin heavy chain (red). Insets show a magnified view of the boxed areas. Bar, 10 µm. (B) The interaction between OCRL1 or INPP5B and the terminal domain of the clathrin heavy chain (CHC TD) or the -adaptin appendage domain (AP-2 -ear) was tested in the yeast two-hybrid system. Growth on high selection indicates an interaction between the proteins. (C) Beads containing immobilised recombinant GST, GST-clathrin terminal domain or GST--adaptin appendage domain were incubated with extracts prepared from HeLa cells expressing GFP-OCRL1 or GFP-INPP5B, and the bound proteins detected by western blotting with antibodies against GFP.