Figure 2.

52 kDa PINK1 interacts with the ERAD machinery. A, PINK1 and Sec61β interaction in WT PINK1 stably-expressing HEK293T cells transfected with Sec61β evidenced by IP using either an anti-PINK1 or anti-Sec61β antibody. *Denotes IgG heavy chain. B, Endogenous PINK1 and Sec61β interaction in SH-SY5Y cells evidenced by IP using an anti-Sec61β antibody. C, WT PINK1 and endogenous gp78 interaction in stably PINK1 expressing HEK293T cells evidenced by co-IP using either an anti-PINK1 or anti-gp78 antibody. D, Ub-PINK1 IB in PINK1 stably-transfected HEK293T cells. Endogenous Sec61β was efficiently knocked down by Sec61β shRNAs. E, Ub-PINK1 IB in PINK1 stably-transfected HEK293T cells. Endogenous gp78 was efficiently knocked down by gp78 shRNA validated by IB. F, Ub-PINK1 IB in Hrd1 deficient HEK293T cells generated by CRISPR that were transfected with WT PINK1. Experiment shown in F was repeated once independently with similar results while those shown in A–E are representative of 3–4 independent experiments performed under identical conditions.