EGF-R is internalized into macropinosomes in HEK-GFP-SNX5 cells. (A) HEK-GFP-SNX5 cells were transfected with EGF-R and 24 h later incubated with Alexa555-conjugated EGF on ice. After washing, cells were incubated at 37°C for 15 min, then fixed and permeabilised and stained with human anti-EEA1 antibody followed by Alexa647-conjugated anti-human IgG. Insert shows overlay of GFP, SNX5 and EEA1. In inset, bar = 5 μm.