Following total RNA extraction using TRIzol reagent (Invitrogen), cDNA was synthesized using Mir-X miRNA First-Strand Synthesis Kit (Takara Holdings Inc., Kyoto, Japan) with extracted total RNA (2 μg). The miRNA expression was measured by qPCR with SYBR Advantage qPCR Premix (Takara) on CFX133a Real-Time PCR Detection System (Bio-Rad Laboratories, Hercules, CA, USA). Relative expression was determined with a U6 control through the 2−ΔΔCt method. The primers used are depicted in Table 1.

Primer Sequences

Abbreviations: miR, microRNA; KDM5C, lysine-specific histone demethylase 5C; PTBP1, polypyrimidine tract-binding protein 1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

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