FIGURE 5.
Membrane and cytosolic FOX activity is partially attenuated in Atp7a KD IEC-6 cells. Membrane and cytosolic proteins were isolated from postconfluent control and Atp7a KD cells. Western blot analysis of protein fractions using a well-established anti-Atp7a antibody (A). Blot shown is representative of 3 independent experiments with similar results. Stained proteins on the blot (shown below) exemplify similar protein loading and efficient transfer onto the membrane. FOX activity by a transferrin-coupled assay is shown in membrane (B) and cytosolic (C) fractions. Different from Ctrl: *P < 0.05, **P < 0.005, ***P < 0.001 (2-factor ANOVA). Values are means ± SDs; n = 6 per group and treatment. Atp7a, Menkes copper-transporting ATPase; Ctrl, control; dA460/dt, change in absorbance over time; FOX, ferroxidase; IEC-6, rat intestinal epithelial; KD, knockdown; M, marker.