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. 2019 Sep 4;39(36):7074–7085. doi: 10.1523/JNEUROSCI.1691-18.2019

Figure 1.

Figure 1.

Ub-PINK1 interacts with VCP. A, Endogenous PINK1 interacting proteins revealed by Coomassie blue-stained SDS-PAGE gel. Endogenous PINK1 and its associated complexes were enriched by IP with anti-PINK1 antibody in HeLa cells. Anti-HA antibody and preimmune IgG (Ctr IgG) were used as controls. Bands labeled 1, 2, and 3 were identified as Ufd2A, VCP, and Npl4, respectively. Individual LC-MS/MS data can be found in Figure 1-1. B, PINK1 and VCP interaction in HEK293T cells cotransfected with WT PINK1 and VCP evidenced by co-IP using either an anti-PINK1 or anti-VCP antibody. *Denotes IgG heavy chain. C, Endogenous PINK1 and VCP interaction in SH-SY5Y cells evidenced by co-IP using anti-VCP-antibody. D, As in B, except that cells were transfected either with WT PINK1 or mutant PINK1K137R. *Denotes IgG heavy chain. E, PINK1 and Ufd2A interaction in HEK293T cells stably-expressing WT PINK1 and transfected with Ufd2A-flag evidenced by IP using an anti-PINK1 antibody. F, Ub-PINK1 IB in WT PINK1 stably-transfected HEK293T cells. Endogenous VCP was efficiently knocked down by VCP shRNAs. G, Ub-PINK1 IB in WT PINK1 stably-transfected HEK293T cells. Endogenous Ufd2A was efficiently knocked down by Ufd2A shRNAs. H, Ub-PINK1 IB in PINK1 stably-transfected HEK293T cells. Endogenous Ufd1 was efficiently knocked down by Ufd1 shRNAs. Data are representative of three independent experiments performed under identical conditions.