Fig. S7.

(A) Time course of α-synuclein deubiquitination by recombinant Usp7 and Usp8. K63-linked ubiquitin chains were conjugated on α-synuclein using purified Nedd4 as the E3. An equal amount of ubiquitinated α-synuclein was incubated with each enzyme (50 nM). Aliquots were taken every 10 min, resolved on SDS/PAGE, and immunoblotted with anti–α-synuclein antibody (C20). (B) SH-SY5Y cells were transduced with Usp8 shRNA or Scr shRNA control. Eight days after transduction, the cells were treated for 24 h with 25 μM chloroquine or 12 h with 1 μΜ lactacystin. α-Synuclein levels were analyzed in total lysates after immunoblotting with the C20 antibody and are presented as the ratio to the actin loading control. Error bars correspond to standard error of the mean.