Mir 23a inhibitor

Hepa1–6 and Hca-P cells were cultured in six-well plates for 12–24 h before transfection. miR-23a mimic, miR-23a inhibitor (antisense oligonucleotide) and scrambled miRNA oligonucleotide pairs were purchased from Ribo-bio (Guangzhou, China). Runx2 siRNAs were designed and synthesized by GenePharma (Shanghai, China). Both cell lines were transfected with 100 nM miR-23a mimic, miR-23a inhibitor, scrambled miRNA or Runx2 siRNA using riboFECT™ CP transfection reagent (Ribo-bio, China) according to the manufacturer’s instructions. The eukaryotic expression plasmid pCMV-Runx2 was obtained from the Karsenty Lab (Columbia University) and was transfected into Hepa1–6 and Hca-P cells using Lipofectamine 2000 (Invitrogen, USA) according to the standard protocol. Cells were collected 48 h after transfection for further assays.

Cells were cultured with RPMI-1640 medium containing 10% FBS overnight, and then 50 nmol/L miR-23a mimic, miR-23a inhibitor and their respective negative control (RiboBio, Guangzhou, China) were transfected into cells using riboFect™ CP Transfection Kit (Ribobio) according to the manufacturer’s instruction, respectively. 24h after transfection, the experiments were performed on the transfected cells.

The synthetic miR-23a mimic, miR-23a inhibitor and CRYAB siRNA were composited by RiboBio (Guangzhou RiboBio Co., Ltd.). The cell lines were transfected with Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA).