ADA2 protein expression and secretion in the homogenous and heterozygous state on a denaturing gel. (A) Immunoblot of whole-cell lysate and supernatants of HEK293T cells transfected with different ADA2 variants in the homozygous state or together with WT ADA2 (heterozygous state). Cells and supernatant were collected 48 h after transfection. The image shown represents three independent experiments. Loading control: B-actin. (B) Quantification of ADA2 protein expression in the whole-cell lysate of HEK293T cells transfected with WT ADA2 or ADA2 variants in homozygous conditions. Bar graphs represent the percentage of ADA2 protein expression relative to WT100% ADA2. (C) Quantification of ADA2 secretion in the supernatant of HEK293T cells transfected with WT ADA2 or ADA2 variants in homozygous conditions. Bar graphs represent the percentage of ADA2 protein expression relative to WT100% ADA2. (D) Quantification of ADA2 secretion in supernatant cotransfected HEK293T cells of ADA2 variants together with WT in heterozygous conditions. Bar graphs represent the percentage of ADA2 secretion relative to WT50% ADA2. (A–D) Each bar represents the mean ± SD from three independent experiments. EV, empty vector. Source data are available for this figure: SourceData F3.