Endosomal trafficking chase of Alexa Fluor 555-conjugated EGF in wild-type and Dync1li1N235Y/N235Y MEFs. A, MEFs were pulsed with EGF for 10 min and chased at 37°C for the times indicated. EGF is shown in red, cytoskeleton (α-tubulin) in green and nuclei in blue (DAPI). Ten cells per genotype per time point were assessed. Scale bar, 30 μm. B, The number of EGF-positive vesicles per cell in wild-type and homozygous MEFs at 0, 20, and 40 min time points. At 0 min wild-type and Dync1li1N235Y/N235Y homozygous mutant cells have similar numbers of vesicles, however, after 20 and 40 min Dync1liN235Y/N235 homozygous mutant cells have significantly more vesicles remaining than wild-type cells.