Figure 7.
Depletion of endogenous PINK1 reduces K63-linked polyubiquitination of IKKγ by parkin. SH-SY5Y cells were transfected with FLAG-tagged IKKγ, Myc-tagged parkin and vehicle (CTL), NT-siRNA or PINK1 siRNA as indicated. In vivo ubiquitination of IKKγ was detected by immunoprecipitation with anti-FLAG antibody followed by immunoblotting using anti-ubiquitin antibody. The levels of PINK1, IKKγ and parkin in the cell lysates were determined by immunoblotting with anti-PINK1, anti-FLAG and anti-Myc antibodies. All experiments were replicated three times with similar results. Ubn-IKKγ, polyubiquitinated IKKγ.