Hypoxia-Reoxygenation Injury Modulation

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.