title: Ribo Seq of shAldh2 and shNC AC16 cells
design: Both shNC and shAldh2 AC16 cells were seeded in 10 cm dishes. Cells were treated with 100 g/mL cycloheximide (CHX) in medium for 3 to 5 minutes after 1 M RSL3 treated for 12 hours to induce ferroptosis at 37C. For Ribo-seq analysis, 1 x 107 cells were rinsed with ice-cold PBS containing 100 g/mL CHX, pelleted (1000g, 10 minutes, 4C), and saved at -80C in nuclease-free tubes. Frozen samples were submitted to APExBIO (Shanghai, China) for library preparation, sequencing, and bioinformatic analysis using two biological replicates following established protocols (Nature. 2024;633:189-197. doi: 10.1038/s41586-024-07781-7).