Figure 13.
Reinforcement of SQSTM1 ameliorates acute lipotoxicity in liver-specific sqstm1 KO mouse livers. sqstm1Alb mice were injected with control vector (Ad-CON)- or SQSTM1 (Ad-SQSTM1)-expressing adenovirus and maintained in a non-fasted state (NF) or fasted overnight and then re-fed a high-carbohydrate, fat-free diet (R). These animals were randomly assigned to 3 groups (5–6 mice in each group). (A) Immunoblot analysis of SQSTM1, KEAP1, p-ULK1(S317), ULK1, ACTB (loading control), nuclear NFE2L2, and LMNB1 (nuclear marker). (B, D) Densitometric analysis of KEAP1 (B) and p-ULK1:ULK1 (D). (C, E-G) qRT-PCR analysis for relative mRNA expression of Keap1 (C), Gsta1 (E), Hmox1 (F), and Nqo1 (G). (H) Liver sections from mice were stained with H&E. CV, central vein. Scale bar: 200 μm. (I) Serum GPT/ALT levels in mice. (J) Images from TUNEL analysis of liver sections. Scale bar: 200 μm. (K) Quantitative analysis of TUNEL-positive cells in liver sections. Data are presented relative to the corresponding value for non-fasted mice and are means ± standard errors for 5–6 mice per group. *p < 0.05, **p < 0.01, and N.S., not significant