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Biological activity of clathrin-box–mutated SMAP1. HeLa cells were transfected with the plasmid expressing the mutated HA-SMAP1 in which amino acid residues 192–196 were replaced with alanine. In A, the cells were incubated with Alexa488-transferrin before fixation to assay transferrin internalization. The cells were fixed and stained with anti-HA antibody alone (A) or together with antibodies specific for AP-2 (B) or CHC (C). Bar, 20 μm.
