Rabbit lc3a b

Homogenates of 6 adult livers from each strain at 16-month-old were used for total protein extraction and kept at −80 °C. Western blot was performed according to the previous reports [71 (link)]. Equal amount protein of each sample was separated in 10% SDS-PAGE gel, transferred onto Polyvinylidene Fluoride (PVDF) membranes (Millipore, Burlington, MA, USA) and blotted with primary antibodies, rabbit Akt (Cell Signaling, 9272S, Boston, MA, USA), rabbit P-Akt (Cell Signaling, 4060S, Boston, MA, USA), rabbit SQSTM1/P62 (MBL, PM045, Woburn, MA, USA), rabbit LC3A/B (Cell Signaling, 4108, Boston, MA, USA), rabbit tP53 (GeneTex, GTX128135, Irvine, CA, USA), rabbit IL-6 (Abcam, ab208113, Cambridge, UK), rabbit Atg5-Atg12 conjugate (Novus biology, NB110-53818SS, Perth, WA, AUS), recombinant anti-Caspase-3 p12 antibody (EPR16888) (Abcam, ab179517, Cambridge, UK) and β-actin (Cell Signaling, 4967S, Boston, MA, USA), separately. Blots were probed with horseradish peroxidase (HRP)-conjugated secondary antibody and visualized using enhanced chemiluminescence reagent (ECL) ((GE Healthcare Bio-Sciences Corp (formerly Amersham Pharmacia Biotech), Piscataway, NJ, USA)) Western blotting detection reagents. NIH software Image J (National Institutes of Health, Rockville, MD, USA) was applied for blot scanning and protein quantifications.