Fig 5.

Nrf2 induction decreases gluconeogenesis-related genes in liver. (A) Expression levels of gluconeogenesis-related genes in livers of 14- to 18-week-old db/db::Keap1^flox/+ (control) or db/db::Keap1^flox/− mice crossed with Nrf2^+/+ or Nrf2^−/− mice (n = 13 to 18). Data are normalized with Hprt. **, P < 0.01; *, P < 0.05. (B) Blood glucose levels in the PTT. Pyruvate (1.5 g/kg of BW) was intraperitoneally administered after 24 h of fasting to 8- to 10-week-old control or db/db::Keap1^flox/− mice (n = 12 to 21). ***, P < 0.001 versus control. (C) Effect of CDDO-Im on G6pc mRNA expression in AML12 cells. After a 24-h incubation with 10 nmol/liter of CDDO-Im, cells were stimulated with 1 μmol/liter of Dex or 1 mmol/liter of dbcAMP for 6 h. Data are represented as G6pc expression levels and normalized with β-actin (n = 4). ***, P < 0.001. (D) Effect of CDDO-Im on G6pc regulatory activities in AML12 cells stably transfected with G6pc-Luc2P. After a 24-h incubation with 10 nmol/liter of CDDO-Im, cells were stimulated with 1 mmol/liter of dbcAMP for 9 h. Data are represented as luciferase activities after normalization to the total cellular protein amounts (n = 5). **, P < 0.01. (E) Role of Nrf2 in G6pc regulatory activities in AML12 cells transiently transfected with G6pc-Luc2P. Data are represented as firefly luciferase activities after normalization to Renilla luciferase activities (n = 6). P < 0.001 (***) and P < 0.01 (**) versus CREB. (F) Immunoblot analysis for CREB in liver and the quantified protein expression levels normalized with α-tubulin of 18-week-old control or db/db::Keap1^flox/− mice (n = 5). ***, P < 0.001. (G) Plasma glucagon concentrations of male 18-week-old Keap1^flox/− or Keap1^flox/+ lean or db/db mice fed ad libitum (n = 3). **, P < 0.01 versus lean mice. Error bars, SEM.