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Further, postmortem alcoholic brains have shown increased TLR2, TLR3, and TLR4 expression in the orbitofrontal cortex, which correlates with BBB integrity loss (Rubio-Araiz et al., 2017)

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When the TSA method was used, the sections were treated with biotinylated goat anti-mouse/rabbit IgG (1:1000 dilution, Vector Laboratories, Burlingame, CA) for 1 h, and then incubated with HRP-conjugated-avidin for 30 min (1:100 dilution in TN buffer, TSA System) at room temperature

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