Livers were washed in ice-cold PBS supplemented with 0.1 mg/mL cycloheximide (Sigma-Aldrich) and minced in lysis buffer (25 mM Tris-HCl [pH 7.5], 250 mM NaCl, 5 mM MgCl2, 0.5 mM PMSF, 0.2 mg/mL heparin [Sigma-Aldrich], 5 mM DTT, 20 U/mL RNasin [Promega], 0.1 mg/mL cycloheximide, 1% Triton X-100, 1× protease inhibitor). Homogenized minced livers were resolved through a 10%–50% sucrose gradient at 200,000g for 2.5 hours at 4°C using a SWT41i rotor (Beckman Brea). Fractions (0.9 mL) were collected and 260 nm absorbance monitored by spectrophotometry. RNA from each fraction was used for qPCR.

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