Fig. 3.
Hepatic neddylation also enhances ETF protein levels in adult mice and prevents fasting-induced steatosis and mortality. (A) IB analysis of NEDD8 and ETF expression in liver tissues of 8-wk-old male mice fasted for 0, 24, or 48 h. (B) IB analysis of ETF expression in adult primary hepatocytes treated with or without 2 μM MLN4924 for 24 h. (C–M) Recombinant AAV-DJ expressing Cre recombinase driven by a strong synthetic CAG promoter was injected into the tail vein of 8-wk-old male Uba3F/F or Nedd8F/F mice and their heterozygous littermates. Six weeks later, the mice were subjected to the following assays: IB (C and E) and IHC (D) analysis of ETF expression in liver tissues of the fed mice; liver weights relative to body weights of the fed mice (F, n = 6 per group); blood parameters before and after a 48-h fast (G–J, n = 6 per group); Nile red staining of liver tissues before and after a 48-h fast (K and L); and survival curves during a 96-h fast (M, n = 6 per group). **P < 0.01. (Scale bars, 50 μm in D; 10 μm in K and L.)