FIG. 1.

In vitro mutagenesis to evaluate ATP8B1 mutation pathogenicity. (A) HEK293T cells were transfected with the indicated ATP8B1 minigene vector and subjected to RNA splicing analysis. (B‐F) CHO‐K1 cells were transfected with WT or mutated pShuttle–ATP8B1‐FLAG, with pShuttle–HA–CDC50A and analyzed for (B,C) ATP8B1 protein expression in whole‐cell lysates, (D,E) cell‐surface fractions, and (F) ATP8B1 flippase activity. ATP8B1‐FLAG expression was quantified. Each bar represents the mean ± SEM of triplicate (C,E) or quadruple (F) determinations. A representative result of two independent experiments is shown. *P < 0.05, **P < 0.01, ***P < 0.001 vs WT. Abbreviations: ATP1A1, adenosine triphosphatase Na+/K+ transporting subunit alpha 1; bp, base pair; BQL, below the limit of quantification; EV, empty vector.