Total RNA was extracted from the CC of control, 4-week BCAS, and 8-week BCAS mice (n = 4/group). miRNA sequencing was performed using the Illumina HiSeq 2500 platform and data analysis was performed by Ribo Bio. Serum miRNAs were extracted from patients using the miRNeasy Serum/Plasma kit (QIAGEN) according to the manufacturer’s manual. Equal amounts of C. elegans miR-39-3p miRNA (miRNeasy Serum/Plasma Spike-In Control, QIAGEN) were added to each sample. Total RNA was extracted from brain tissue or cultured cells using TRIzol (Invitrogen) followed by cDNA synthesis by a RevertAid First-Strand cDNA Synthesis Kit (Thermo Fisher Scientific). TaqMan probe– or SYBR Green–based quantitative PCR was conducted on a Stratagene Mx3000P real-time PCR system (Agilent Technologies). Specific stem-loop primers were used for reverse transcription of miRNAs (Supplemental Table 4). The primer pairs are listed in Supplemental Table 5.

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