Cell strainer

At the later stage, three 12-month-old male NORAD−/− mice and six 12-month-old male wild-type mice were randomly selected. Three NORAD−/− mice and three wild-type mice received tail vein injections of NaIO₃ (Sigma-Aldrich, China) (30mg/kg). Eight days later, the heart, liver, lungs, kidneys, brains, and eyes (one eye is used for RT-qPCR experiments later) were dissected from mice. All organs were filtered into a single-cell solution by a cell strainer (Biosharp, China), then washed five times with PBS for the determination of the ROS experiment. The remaining eyes were employed for the detection of AMD markers such as C3, ICAM-1, APP, APOE, and CRYAA (for the primer sequence see Supplementary Table 1).

Newborn mice aged 1–3 days were subjected to gentle meningeal dissection using ophthalmic forceps, scissors, and other tools. Subsequently, the brain tissue was carefully peeled off and placed in a 0.25% trypsin solution (Gibco, 15050065). The tissue was then incubated in a cell culture incubator at 37 °C for 5 min. After the incubation period, the trypsin reaction was stopped by adding complete culture medium, and the cell suspension was filtered through a cell strainer (Biosharp). The filtrate was collected and subjected to centrifugation at 1000g for 10 min. The supernatant was discarded, and the cells were resuspended in complete culture medium. The cells were then seeded in T-75 cell culture flasks containing 10% fetal bovine serum (FBS) (Sigma, F8318)-F12/DMEM culture medium and were subsequently maintained through medium changes and passaging for experimental use.

Hepatocytes were obtained from the liver tissue of mandarin fish. The liver tissue was cut into small pieces and digested with trypsin (Gibco, USA). Tissue was dispersed into cells through cell strainer (Biosharp, China). Red cells were lysed with red cell lysis buffer (Biosharp, China). Cells were cultured with M199 (10% fetal bovine serum, penicillin-streptomycin solution 1‰) (Gibco, USA). The inhibitor of SET1DB chaetocin (17 (link)) (Selleck, USA) was used to treat the cells at a concentration of 3 × 10⁻⁵ mol/L for 17 h, and then the levels of H3K4me3 and pepck mRNA expression were examined with six biological replicates and three technical replicates.