Fig. 5.
Inhibition of TBK1 suppresses p62 phosphorylation and accumulation in livers of obese mice. (A,B) Four-month-old C57BL/6 male mice kept on HFD for 2 months were subjected to daily administration of phosphate-buffered saline (Con, n = 7) or BX795 (25 mg/kg body weight, i.p.; n = 6) for 10 days. LFD-kept mice (n = 5) of the same age were used as a negative control. (C,D) Five-month-old littermate-controlled Tbk1F/F (n=16) and Albumin-Cre/Tbk1F/F (n=16) male mice of C57BL/6 background, kept on HFD for 3 months, were analyzed. (A,C) Livers were subjected to solubility fractionation. 1% Triton X-100-soluble and -insoluble fractions were analyzed by immunoblotting and immunoblot quantification. (B,D) Liver sections were subjected to p62 immunostaining. Hematoxylin counterstaining was used to visualize nuclei. Boxed areas in top panels are magnified in bottom panels. Scale bars, 200 μm. All data are shown as mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001 (Student’s t-test). Arrowheads indicate the exact or nearest position of the protein molecular weight markers (kD).