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International Journal of Environmental Research and Public Health , 16(1), 146

Immunohistochemistry was performed as follows: free-floating VTA brain sections were incubated 1 h at 4 C in a blocking solution of phosphate-buffered saline (PBS) containing 3% bovine serum albumin (BSA, Sigma A4503) and 0.2% Triton X-100 and then incubated overnight at 4 C with a mouse anti-tyrosine hydroxylase antibody (TH, Sigma, T1299) at 1:500 dilution in PBS containing 1.5% BSA and 0.2% Triton X-100 (see supplementary Table 1)

900 was selected in previous investigations based on simulation studies as the number that generated sufficiently precise estimates for a wide range of outcomes