Mir 15a 5p mimic

The miR-15a-5p mimic, mimic negative control (NC), miR-15a-5p inhibitor, inhibitor NC, YAP1 overexpression plasmid (pcDNA-YAP1) and pcDNA-vector were all provided by Guangzhou RiboBio Co., Ltd. When C-33A and SiHa cells (5×10⁵ cells/well) in 6-well plates grew to ~80% confluence, miR-15a-5p mimic (20 nM), mimic NC (20 nM), miR-15a-5p inhibitor (20 nM), inhibitor NC (20 nM), pcDNA-YAP1 (2 µg) or pcDNA-vector (2 µg) were transfected into cells at 37°C for 24 h using Lipofectamine^® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.). The sequences were as follows: miR-15a-5p mimic, 5′-UAG CAG CAC AUA AUG GUU UGU G-3′; mimic NC, 5′-UUC UCC GAA CGU GUC ACG UTT-3′; miR-15a-5p inhibitor, 5′-CAC AAA CCA UUA UGU GCU GCU A-3′; and inhibitor NC, 5′-CAG UAC UUU UGU GUA GUA CAA-3′.
In addition, small interfering RNA targeting YAP1 (si-YAP1) and the negative control targeting a non-specific sequence (si-Scramble) were provided by Thermo Fisher Scientific, Inc. SiHa and C-33A cells were transfected with the siRNAs (100 nmol/l) using Lipofectamine 2000 (Invitrogen; Thermo Fisher Scientific, Inc.). The sequences of si-YAP1 and si-Scramble were as follows: si-YAP1, 5′-CTC AGG ATG GAG AAA TTT A-3′; and si-Scramble, 5′-TTC TCC GAA CGT GTC ACG T-3′. At 24 h post-transfection, the cells were harvested for further analysis, and the inhibition efficiency was deter-mined by western blotting.