Mice were sacrificed three days after MPTP treatment and the SN was dissected out. SN lysates containing equal amounts of protein were loaded in each lane and separated in a 10-15% SDS-polyacrylamide gel, as described previously (Jin et al., 2011 (link)). Proteins were transferred to a nitrocellulose membrane and nonspecific binding sites were blocked with Licor Odyssey blocking buffer. The membranes were then incubated with different primary antibodies such as anti-IBA-1 (Abcam), anti-GFAP (EMD Millipore), anti-iNOS (Santa Cruz), anti-gp91phox (Abcam), anti-3NT (EMD Millipore) and anti-4HNE (R & D Systems). Next, membranes were incubated with one of the following secondary antibodies: Alexa Fluor 680 goat anti-mouse, Alexa Fluor 680 donkey anti-goat (Invitrogen) or IR dye 800 donkey anti-rabbit (Rockland). To confirm equal protein loading, blots were reprobed with a β-actin antibody (Sigma; 1:10000 dilution). Western blot images were captured with a Licor Odyssey machine (Licor, NE), and the bands were quantified using NIH ImageJ software.