Figure 5: Gluconeogenesis is unaltered in SIK2/AMPKα1/AMPKα2 triple knockout hepatocytes.
From: The LKB1-salt-inducible kinase pathway functions as a key gluconeogenic suppressor in the liver
(a) Glucose production was assessed by measuring glucose in the culture medium of primary hepatocytes isolated from control and liver SIK2/AMPKα1/AMPKα2 triple KO (SIK2/AMPK TKO) male mice (10-week-old) 8 h after treatment with or without 100 μM Bt2-cAMP or 1 μM glucagon. Glucose production was normalized to protein content and expressed as a percentage of glucose production by control hepatocytes incubated in the absence of Bt2-cAMP or glucagon. (b) Immunoblotting of SIK2, AMPKα, phospho (P)-CRTC2/CRTC2, PEPCK and GAPDH in primary hepatocytes isolated from control and liver SIK2/AMPKα1/AMPKα2 triple KO (SIK2/AMPK TKO) mice and treated for 8 h with or without 100 μM Bt2-cAMP or 1 μM glucagon. (c) Immunoblotting of SIK2, AMPKα, P-AMPKα, ACC, P-ACC, P-CRTC2/CRTC2 and GAPDH in primary hepatocytes isolated from control and liver SIK2/AMPKα1/AMPKα2 triple KO (SIK2/AMPK TKO) mice and treated for 8 h with or without 100 μM Bt2-cAMP and with or without 0.25, 0.5 or 1 mM metformin as indicated. (d) Gluconeogenic gene expression was measured by quantitative reverse transcription–PCR in primary hepatocytes isolated from control (white bars) and liver SIK2/AMPK TKO (black bars) mice and treated for 8 h with or without 100 μM Bt2-cAMP or 1 μM glucagon. Transcript levels in control cells were assigned an arbitrary value of 1.0 for comparison. All values are presented as mean±s.e.m. Results are representative of at least three independent experiments.
