Figure 6. COMMD1 diminishes Parkin C289G aggregation.

A. HEK293T cells were transfected with Parkin WT and C289G constructs alone, or in combination with HA-COMMD1. Supernatant (Sup) and pellet fractions were prepared as described in Experimental procedures. Immunoblotting was performed using indicated antibodies. Graphs represent relative fraction of insoluble Parkin proteins in absence or presence of exogenous COMMD1 (fractions quantified from both Parkin as well as Flag immunoblots). B. HEK293T cells were transient transfected with GST or COMMD1-GST alone in combination with Flag-Parkin constructs (WT and C289G). Cells were lysed, and GST fusion proteins were precipitated as described under Figure 1A. Immunoblotting was performed using indicated antibodies. Densitometric quantification of interaction strength between COMMD1 and Parkin WT versus C289G mutant, normalized for total Parkin expression (input). Binding of COMMD1 to Parkin WT was set at 1.