Figure 1

Association of HPS3 with clathrin. (a) Immunoprecipitation of clathrin with HPS3 antibodies. Western blots of normal (NL) and HPS3-null (HPS3) melanocyte lysates treated with clathrin heavy chain antibodies. Equal amounts of clathrin were detected in both lysates [left panel]. The lysates were immunoprecipitated with HPS3 antibodies, electrophoresed, and immunoblotted using clathrin heavy chain antibodies. HPS3 antibodies immunoprecipitated clathrin only in the normal and not in the HPS3-null melanocyte lysates [right panel]. (b) Amino acid sequences of the predicted clathrin-binding domain (residues 172–176 of human HPS3, shown in blue) in the human, mouse and rat HPS3 proteins, and the surrounding amino acid sequences. (c) Confocal immunofluorescence microscopy of representative normal melanocytes electroporated with GFP-HPS3 [A] and GFP-HPS3-delCBD [B] (in green) and co-stained with antibodies to clathrin (in red). GFP-HPS3 partially co-localized with clathrin on small vesicles (arrowheads) [A], but GFP-HPS3-delCBD did not [B]. (Bar = 20 μm).