Fig. 5.

Caveolin-1 (Cav-1) is required for ATP7A protein stabilization and prevents proteosomal degradation. A: Cav-1 wild-type (WT) and Cav-1^−/− [knockout (KO)] mouse fibroblasts were lysed and immunoprecipitated (IP) with anti-ATP7A, followed by immunoblotting (IB) with an anti-ubiquitin antibody. Right: averaged data for ATP7A ubiquitination (n = 3). B: mouse fibroblast cells were incubated with an inhibitor of the proteasome MG132 (20 μmol/L) for 24 h or an inhibitor of the lysosome chloroquine (100 μmol/L) for 24 h, and the protein expression of ATP7A was determined by Western blot (n = 3). C: membrane fractionation of mouse aorta. Equivolume fractions isolated from the top (fraction 1) to the bottom (fraction 13) were immunoblotted with antibodies as indicated. D: IP of Cav-1 using an anti-Cav-1 antibody in aortic lysates, followed by IB with an ATP7A antibody (n = 3). Results are presented as means ± SE. *P < 0.05. ATP7a, Menkes ATPase, copper-transporting P-type ATPase.