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The samples were then permeabilized in 0.5% Triton X-100 for 3 min and blocked with 3% goat serum albumin for 30 min prior to incubation with a primary antibody, namely, anti--synuclein, mouse anti-TH, SIGMAR1 Ab (Affinity Biosciences, Cat.#: DF7363) and anti-NeuN, at dilutions of 1:100 overnight at 4 C