Total RNA fractions were isolated from the mouse frozen tissue (n =30 adolescents and n =26 adults) with the miRNeasy Micro kit protocol (QIAGEN). All RNA samples were determined to have 260/280 and 260/230 values ≥1.8, using the Nanodrop 1000 system (Thermo Scientific). Reverse transcription for Dcc mRNA was performed using High-Capacity cDNA Reverse Transcription kit (Applied Biosystems). qPCR (Reynolds et al., 2015; Torres-Berrío et al., 2017) using TaqMan assay (Applied Biosystems) was conducted with an Applied Biosystems QuantStudio 6 Flex Real-Time PCR system. Data for Dcc expression were analyzed by using the relative quantification method and Gapdh was used as reference gene. Real-time PCR was run in technical triplicates.

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