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The pcDNA empty vector (NC, 5 μg/mL), pcDNA-MIR497HG (MIR497HG, 5 μg/mL), siRNA normal control (si-NC, 50 nM), siRNAs against MIR497HG (si-MIR497HG, 50 nM), miRNA control (miR-NC, 50 nM), miR-29b-3p mimics (miR-29b-3p, 50 nM), and miR-29b-3p inhibitors (miR-in, 100 nM) were supplied by GenePharma Co., Ltd. (Shanghai, China). BV2 and HT-22 cells, inoculated into 24-well plates with a density of 3 × 10⁵ cells/well, were incubated with 5% CO₂ at 37°C for 24 hours and then subjected to cell transfection. Lipofectamine® 3000 (Invitrogen; Thermo Fisher Scientific, Inc.) was taken to transfect the above substances into BV2 and HT-22 cells as per the supplier's instructions. qRT-PCR confirmed the transfection efficiency. The cells were incubated with 5% CO₂ at 37°C for 24 hours in preparation for further analysis. Following 24 hours' transfection, CGA and OGD were adopted to treat BV2 and HT-22 cells [12 (link)].

MiR-146a-5p mimics (miR-146a-5p), mimics negative control (miR-NC), miR-146a-5p inhibitors (miR-146a-5p-in), inhibitors negative control (miR-in), empty plasmid and TRAF6 overexpression plasmid were available from GenePharma (Shanghai, China). PAF-induced HSAECs were transfected by lipofectamine^TM 3000 (Invitrogen, Carlsbad, CA, USA), and 48 h later, the transfection efficacy was accordingly examined by quantitative real-time polymerase-chain reaction (qRT-PCR).

We ordered HCC cell lines (HCCLM3 and Huh-7) and a hepatocellular cell line L-O2 from the American Type Culture Collection (Manassas, VA, USA). Dulbecco’s Modified Eagle’s medium (DMEM) comprising 10% fetal bovine serum (FBS, Gibco, Thermo Scientific, Shanghai, China), 100 U/mL penicillin, and 100 μg/mL streptomycin (Gibco, Thermo Scientific, Shanghai, China) was applied for cell growth. Then, cell culture was performed at 37°C with 5% CO₂.
We obtained miR-142-3p mimics, negative controls (miR-NC), miR-142-3p inhibitors and their negative controls (miR-in) from GenePharma (Shanghai, China). Then, the pcDNA3.1-PIK3CG expression vector was built by embedding the overall length of PIK3CG cDNA into the backbone of the pcDNA3.1 vector. Next, the transfection was conducted by utilizing lipofectamine 2000 (Invitrogen, Grand Island, NY, USA) as guided by the manufacturer. In short, the miRNA (50 nM) or plasmid (100 µg) undergoing dilution in 100 µL of OPTI-MEM (Thermo Scientific, Shanghai, China) was subjected to mix with 10 µL of lipofectamine for 15-minute incubation at room temperature (RT). Finally, 48 hours of culture was carried out after adding the mixture to the cells for further analysis [19 (link)].