Figure 3. Gluconeogenesis remains unaltered in SIK2-knockout hepatocytes.

(a) Immunoblotting of SIK2, CRTC2, PEPCK and GAPDH in primary hepatocytes isolated from control and liver SIK2-KO male mice (10-week-old) that had been treated for 8 h with or without 100 μM Bt₂-cAMP. (b) Gluconeogenic gene expression measured by quantitative reverse transcription–PCR in primary hepatocytes isolated from control (white bars) and liver SIK2-KO (black bars) mice and treated for 8 h with or without 100 μM Bt₂-cAMP. Transcript levels in control cells were assigned an arbitrary value of 1.0 for comparison. (c) Glucose production was measured in the culture medium of primary hepatocytes isolated from control and liver SIK2-KO mice 4, 8, 12 or 24 h after treatment with or without 100 μM Bt₂-cAMP as described in Methods. Glucose production was normalized to protein content and expressed as a percentage of glucose production by control hepatocytes incubated for 4 h in the absence of Bt₂-cAMP. All values are presented as mean±s.e.m., n=3.