Ribosome profiling of Atg12 KO MEFs
Ribosome profiling experiments were performed using the ARTseq Ribosome profiling kit (Epicentre). Briefly, immortalized Atg12f/f or Atg12KO MEFs were maintained in control media (DMEM + 10% fetal bovine serum) or starved in HBSS for 2 hours. Following the starvation period, cycloheximide made fresh to 50mg/ml in Ethanol for each experiment was added at a final concentration of 100ug/ml.  Then, cells were collected in PBS and preparation of the samples was performed according the ARTseq Ribosome profiling kit manufacturers instructions. RNA extraction by Trizol LS (Ambion), rRNA depletion via RiboZero Gold (Epicentre), and quality and quantity of small RNA and DNA assayed using Agilent High Sensitivity Small RNA kit and DNA kit respectively (Agilent).