Mir 129 5p mimic

The human miR-129-5p gene was PCR-amplified from genomic DNA and cloned into a pMSCV-puro retroviral vector. The TEAD Luciferase Reporting system was purchased from lifeome (Oceanside, CA, USA). We purchased miR-129-5p mimic, miR-129-5p antagonist (antagomiR-129-5p), and controls from RiboBio (Guangzhou, China). Transfection of the plasmids, siRNAs, miR-129-5p mimic, and antagomiR-129-5p were performed using Lipofectamine 2000 (Invitrogen) according to the manufacturer's instructions.

Mouse HSCs in the logarithmic growth phase were cultured in DMEM containing 10% FBS. When cell confluence reached 40–60%, HSCs (5 × 10³ cells/well) were transfected with the following plasmids according to the instructions of the Lipofectamine 2000 kit (11668-027, Invitrogen Inc., Carlsbad, CA, United States): pcDNA3-shRNA-NC, pcDNA3, pcDNA3-shPEG3, miR-NC, miR-129-5p mimic, pcDNA3-NEAT1, pcDNA3-PEG3, or their respective NCs. Besides, cells were treated with dimethyl sulfoxide (DMSO) or pyrrolidine dithiocarbamate (PDTC, an inhibitor of the NF-κB signaling pathway) (2 mmol/l for 24 h). All plasmids and miR-129-5p mimic were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).