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. 2014 Apr 1;9(4):e92408. doi: 10.1371/journal.pone.0092408

Figure 7. Aggregation of polyglutamine proteins is unaffected by COMMD1.

Figure 7

A. HEK293T cells were transfected with eYFP-HttQ74 and eYFP-HttQ119 alone, or in co-expression with HA-COMMD1 or V5-DNAJB6 prior to lysis and detection of polyQ HMW complexes using SDS-PAGE. HMW aggregating complexes were trapped and detected in the stacking gel. IB was performed using indicated antibodies. Samples were analysed on the same membranes. Graph represents the relative fraction of insoluble HttQ74 and Q119 HMW species under EV conditions or upon co-expression of COMMD1 or DNAJB6. Insoluble Htt HMW species present in EV conditions were set at 1. B. Filter trap analysis of HEK293T cells transfected with the polyQ proteins eYFP-HttQ74 and eYFP-HttQ119, in co-expression with EV or HA-COMMD1. Samples were prepared as described under Experimental procedures and diluted as indicated. Immunoblotting was performed antibodies directed against the eYFP-tag. Densitometric quantification of insoluble fraction of HttQ74 and HttQ119, in absence or presence of exogenous COMMD1 (5x dilution). EV conditions were set at 1. p  =  0.0425, indicates a significant decrease in insoluble HttQ74 aggregates mediated by COMMD1. n.s.  =  non-significant.