As previously (Torres-Berrío et al., 2017; Cuesta et al., 2020a), protein samples (20 μg, n =25 adolescents and n =24 adults) were separated on a 10% SDS-PAGE and transferred to a PVDF membrane which was incubated overnight at 4°C with antibodies against Netrin-1 (Abcam, catalog #126729) and α-tubulin (Cell Signaling, catalog #2144S) for loading control. Four biological samples were included per experimental condition. Since Netrin-1 is a secreted protein which can be produced remotely (Brignani et al., 2020), we routinely quantify protein rather than mRNA levels.

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