Posttranscriptional reprogramming controls MASLD progression through chronic ER stress adaptation (Riboseq)
Posttranscriptional reprogramming controls MASLD progression through chronic ER stress adaptation (Riboseq)
A piece of aproximately 30 mm3 of snap frozen murine liver was homogenized in lysis buffer [20 mM Tris-HCl pH 7.8, 10 mM MgCl2, 100 mM KCl, 1% Triton X-100, 2 mM DTT, 100 μg/ml CHX, complete protease inhibitors] using a bead beating grinder.  Lysates were passed through a 26G needle (BD bioscience) for further homogenization, and centrifuged at 4°C 1300xg for 10 min. Supernatants were collected and used to perform Ribosome profiling (RP) and total mRNA isolation, as previously described in Slobodin et al. 2017. Male C57BL/6J mice were housed under standard conditions. At 6 weeks of age, mice were randomly assigned to either a high-fat diet (HFD) or a normal diet (ND) and fed HFD or ND for 30 weeks when the experiments were perfomed. A piece of aproximately 30 mm3 of snap frozen murine liver was homogenized in lysis buffer [20 mM Tris-HCl pH 7.8, 10 mM MgCl2, 100 mM KCl, 1% Triton X-100, 2 mM DTT, 100 μg/ml CHX, complete protease inhibitors] using a bead beating grinder.  Lysates were passed through a 26G needle (BD bioscience) for further homogenization, and centrifuged at 4°C 1300xg for 10 min. Supernatants were collected and used to perform Ribosome profiling (RP) and total mRNA isolation, as previously described in Slobodin et al. 2017. as previously described in Slobodin et al. 2017