SiRNA silencing of AP complexes 1 and 2 (AP-1 and AP-2). HEK293T cells were treated with 200 nM of AP-1 siRNA (sigma 1 and gamma subunits) or AP-2 siRNA (sigma 2 and alpha subunits) or non-target siRNA, by Lipofectamine 2000 (Invitrogen). After 48 h, the cells were transfected with Venus-tagged ATP7A. (A and B) Expression of AP-1 in non-target siRNA and AP-1 siRNA-transfected cells, illustrated by representative western blots of AP-1 gamma subunit. (B and C) Expression of AP-2 in non-target siRNA and AP-2 siRNA-transfected cells, illustrated by representative western blots of AP-2 alpha subunit; (D) Summary of densitometric analyses of five western blots. Error bars = SEM. KD, knock down.