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Fig. S1. FeCl2 treatment inhibits proteasomes. (A) Immunoblotting of cell lysates with anti-ubiquitin antibody (Mab 1510, Chemicon). FeCl2-treated samples displayed more high-molecular-weight (HMW) proteins with ubiquitin (ubiq) immunoreactivities. The arrow indicates ubiquitin monomer. (B) Quantitative analysis of ubiquitin-immunoreactive proteins of molecular weight > 75 kDa. Student?s t-test, N = 6, *P < 0.05. (C) Proteasomal activity assay using fluorogenic peptides as substrate. Chymotrypsine-like activity was lower in Fe(+) than Fe(?) controls, and higher in Syn(+) than Syn(?) cells without the FeCl2 treatment, N = 6, *P < 0.05. The specificity of activity assay was verified by analysis of lysates in the presence of the proteasome inhibitor, epoxomicin.

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