WT_mRNAseq_3
A total of 10^7 cells were prepared for cell lysis. Then, the cell medium was discarded, and the cells were washed twice with ice-cold PBS containing 100 g/mL cycloheximide (Adooq, 66-81-9). 100 mg tissue were pulverized manually under liquid nitrogen and treated by ice-cold PBS containing 100 g/mL cycloheximide. Ribosome profiling was performed using Epi Ribosome Profiling Kit (Epibiotek, R1814). Subsquently, RPFs (ribosome-protected RNA fragments) was extracted using RNA clean&ConcentratorTM-5 kit (ZYMO, R1016). EpiTM RiboRNA Depletion Kit (Human/Mouse/Rat) (Epibiotek, R1805) was used for rRNA depletion. Sequencing libraries were constructed using QIAseq miRNA Library kit (QIAGEN, 1103679).