Mir 29a 3p mimics

Human PTC cell lines K1 and TPC-1, the human thyroid follicular epithelial cell line, Nthy-ori3-1, and HEK 293 T cells were purchased from the Shanghai Cell Bank (Shanghai, China). All cell lines were grown in RPMI 1640 medium (BI, Jerusalem, Israel), supplemented with 10% fetal bovine serum (BI), and incubated at 37°C in a humidified incubator containing 5% CO₂. miR-29a-3p mimics, control mimics, specific small interfering RNA (siRNA) for OTUB2, and control siRNA were synthesized by RiboBio Inc (Guangzhou, China) according to the published work.19 (link),20 (link) miRNA mimics and siRNAs with final concentration at 50 nM were transfected into PTC cells using Lipofectamine™ 2000 (Thermo Fisher Scientific, Waltham, MA, USA) according to manufacturer’s instructions. Briefly, nucleic acids, which includes the plasmids and siRNAs, and the Lipofectamine reagent were diluted with RPMI 1640 medium, for 5 minutes. Then the mediums containing nucleic acids and Lipofectamine reagent were mixed and incubated at room temperature to form liposomes. After 15 minutes, each mixture was added into the corresponding culture medium and 6 hours later, the culture medium was removed and replaced with fresh medium.