Fig. 5.

Retromer recycles CTR1 back to the cell surface in response to a reduction in extracellular copper concentration. CTR1 surface levels were analysed under copper-induced (200μM CuCl₂ for 1 h) or copper washout (recovery in control medium for 1 h) conditions in control and VPS35-depleted cells. At the indicated time points cells were surface biotinylated and streptavidin–agarose used to capture biotinylated membrane proteins. The surface abundance of CTR1 and the loading control N-cadherin were examined by immunoblotting. The graph represents the means±s.e.m.; n=3 independent experiments. **P<0.01 (unpaired Student's t-test).