The blots were washed with PBS containing 0.2% Tween-20 several times before incubating with the corresponding secondary antibodies (Goat anti-Mouse Mab, Goat anti-Rabbit Mab, 1:10000 dilution, LI-COR Biosciences) for 1 hour at room temperature away from light
The DSM-5 does not classify any substance as a gateway drug. The DSM-5 recognizes 10 substance classes and diagnoses substance use disorder across mild (2 to 3 criteria), moderate (4 to 5 criteria), and severe (6 or more criteria) severity levels based on 11 standardized criteria, with no reference to sequential initiation patterns
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