*4.11. Western Blot*

Equal amounts of total protein extracts were electrophoretically resolved on 8% SDS-PAGE. Resolved proteins were then transferred onto nitrocellulose membranes and blocked with a 5% milk solution in Tris-buffered saline (TBS). Then, membranes were incubated with primary antibody rabbit polyclonal TH (#ab112, Abcam), overnight at 4 ◦C. After washing with TBS/0.2% Tween 20 (TBS-T), membranes were incubated with secondary goat anti-rabbit IgG antibody conjugated with horseradish peroxidase (Bio-Rad Laboratories) for 2 h at room temperature. Membranes were processed for protein detection using ImmobilonTM Western (Millipore). β-actin (AC-15) (#A5441, Sigma-Aldrich) was used as loading control. Densitometric analysis was performed using the Image Lab Software version 5.1 Beta (Bio-Rad).
