Figure 6.

Trehalose upregulation of hepatocyte UCP1 is PPARGC1A- and TFEB-dependent. (A) Ucp1 mRNA accumulation in AML12 murine hepatocytes and primary murine hepatocytes treated with or without trehalose for 24 h (n = 3–9 cell cultures per group) and in liver from mice treated with or without oral trehalose (3% water, 5 days). (B) UCP1 immunoblot analysis in untreated iBAT or liver tissue from mice treated with or without trehalose (upper panels). UCP1 immunoblot analysis in untreated iBAT or primary hepatocytes treated with or without trehalose (24 h; lower panels). (C) Ucp1 mRNA accumulation in primary hepatocytes treated with or without trehalose in the presence or absence of 5 mM pyruvate. (D) Ucp1 mRNA accumulation in primary hepatocytes treated with or without trehalose in the presence or absence of scrambled or Ppargc1a-directed ASO. (E) Ucp1 mRNA accumulation in primary hepatocytes treated with or without trehalose in the presence or absence of DMSO or LY2874455. (F) Ucp1 upregulation in primary hepatocytes incubated with low serum and low glucose (‘Starve’, 24 h) or regular growth media (‘Control’, 24 h) in the presence of scrambled or Ppargc1a- or Fgf21-directed ASO. (G) Ucp1 mRNA accumulation in primary hepatocytes treated with or without trehalose in the presence or absence of scrambled or Tfeb-directed siRNA (left). Tfeb mRNA abundance in Tfeb siRNA-treated hepatocytes (24 h; right). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 versus control by Student’s T-test with Bonferroni-Dunn post-hoc correction for multiple tests.