U6 small nuclear rna rnu6b

Total RNA was isolated from tissue samples and cell lines using TRIzol reagent (Invitrogen) according to the manufacturer’s protocol. The levels of mature miR-141 (miR-141) and precursor miR-141 (pre-miR-141) were evaluated using the miDETECT A Track miRNA qRT-PCR Kit (RiBoBio). The U6 small nuclear RNA (RNU6B) (RiBoBio) was used for normalization. The expression of primary miR-141 transcript (pri-miR-141), c-Myc and BRD7 was measured by qRT-PCR according to the instructions of the SYBR Premix Ex Taq (TaKaRa, Dalian, China). The GAPDH mRNA level was used for normalization. The relative expression ratio was calculated using the 2^−ΔΔCT method. The primers for miR-141, pre-miR-141, U6, pri-miR-141, c-Myc, BRD7, and GAPDH were described previously [23 (link)]. PCRs of each sample were conducted at least in triplicate.

Total RNA was isolated from tissue samples and cell lines using TRIzol reagent (Invitrogen) according to the manufacturer's protocol. The expression level of mature miR-141 and pre-miR-141 was evaluated using the miDETECT A Track miRNA qRT-PCR Kit (RiBoBio) following the manufacturer's protocol. The U6 small nuclear RNA (RNU6B) (RiBoBio) was used for normalization. The expression level of pri-miR-141 was measured by qRT-PCR according to the instructions of the SYBR Premix Ex Taq (TaKaRa, Dalian, China). The GAPDH mRNA level was used for normalization. The relative expression ratio of mature miR-141, pre-miR-141 and pri-miR-141 was calculated using the 2^−ΔΔCT method. The primer for mature miR-141 was 5′-TAACACTGTCTGGTAAAGATGG-3′ (forward). Primer for pre-miR-141 was 5′-TTGGATGGTCTAATTGTGAAGCTCC-3′ (forward). Primer pairs for U6 were 5′-ATTGGAACGATACAGAGAAGATT-3′ (forward) and 5′-GGAACGCTTCACGAATTTG-3′ (reverse). Primer pairs for pri-miR-141 were 5′-AGACCTCACCTGGCCTGTGGCC-3′ (forward) and 5′-GAACCCACCCGGGAGCCATCTT-3′ (reverse). Primer pairs for GAPDH were 5′-CGAGATCCCTCCAAAATCAA-3′ (forward) and 5′-TTCACACCCATGACGAACAT-3′ (reverse). PCRs of each sample were conducted in triplicate.