Si mettl14

Human umbilical vein endothelial cell line (HUVECs, EAhY926) purchased from the ATCC was used in the experiment. The cells were cultured with DMEM medium supplemented with FBS (10%). 100 μL/mL of ox-LDL was added in the medium and incubated with target cells for 12 h or 24 h to establish cell models for atherosclerosis. si-NC and si-METTL14 purchased from GenePharma (Shanghai, China) were transfected into HUVECs by Lipofectamine 2000 (Invitrogen, CA, USA). pcDNA 3.1-p65 or pcDNA 3.1-NC adenoviral vectors obtained from HanBio Technology Co. Ltd. (Shanghai, China) were transfected into HUVECs for the overexpression of p65 or used as a control in the following experiment.

Small interfering RNAs of METTL14 (si-METTL14), APPL1 (si-APPL1), and their negative control (si-NC) were synthesized and obtained from GenePharma (Shanghai, China). miR-146a-5p mimics and mimics NC were also provided by GenePharma. As prescribed by the manufacturer’s protocol, Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) was employed to transfect the above vectors. In brief, HL-1 cells were cultured until 60–80% confluence and transfected with vectors. After transfection, HL-1 cells were subjected to H/R treatment. During H/R treatment, 1 × 10⁵ cells were seeded into the 6-well plates and were cultured in an anoxic chamber (94% N₂, 5% CO₂, and 1% O₂) for 16 h, followed by 6 h culture under a normal oxygen environment of 5% CO₂ for reoxygenation [33 (link), 34 (link)]. Cells in the control group were cultured under conventional conditions.