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16E5 inhibits endosome acidification and trafficking. Trafficking and acidification of endosomes in unfixed EGF-starved HFKs expressing 16E5 (or empty pLXSN vector) after labeling with Alexa Fluor 488-EGF and pHrodo-EGF at 4°C and warming to 37°C for up to 70 min was examined. Where indicated, 0.33 μM bafilomycin A1 was added to cells as they were shifted to 37°C (+BfA). Scale bar, 20 μm.
