Functional characterization of variants in ABCA1. A: Western blot analysis showing normal protein expression of wild-type and mutant ABCA1. The two ABCA1 variants, as well as wild-type ABCA1, were generated in vitro and expressed in HEK293 cells. Cell lysates were probed with antibodies to ABCA1 and calnexin (as loading control). Neg, negative control. B: Immunofluorescence results showing normal subcellular localization of wild-type, W590L, and R2200* ABCA1 proteins. C: HEK293 cells were transiently transfected with wild-type, W590L, and R2200* ABCA1, and cholesterol efflux measured in the presence or absence of ApoA-I. # Indicates a statistically significant difference compared with mock transfected and * indicates a statistically significant difference compared with wild-type ABCA1 at P < 0.05 using a one-way ANOVA.