Cell culture
COS-7 cells were used to express EFEMP1 protein. The COS-7 cells were seeded in 6-well plates at a concentration of 100,000 cells per mL and allowed to grow in DMEM + 10% FBS condition media. The cells were transfected with expression clones encoding wild-type or mutantEFEMP1 cDNA using Lipofectamine 3000 (Invitrogen, Carlsbad, CA) 24 hours post-seeding at 80-90% confluence.