Figure 7: Effect of HG-9-91-01 in AMPK- or LKB1-knockout primary hepatocytes.
From: The LKB1-salt-inducible kinase pathway functions as a key gluconeogenic suppressor in the liver
(a) Primary hepatocytes from control (AMPKα1lox/loxα2lox/lox) and liver-specific AMPKα1/α2 knockout (AMPK KO) male mice (10-week-old) were treated with the indicated amount of HG-9-91-01 for 8 h prior to cell lysis. Immunoblot analysis was conducted with the indicated antibodies and glucose was measured in the culture medium. Glucose production was normalized to total protein content and presented as percentage of glucose production by non-treated hepatocytes. Data is presented as mean±s.d., n=3. *P<0.01 basal versus 1 μM HG-9-91-01 treatment in both genotypes and #P<0.001 basal versus 4 μM HG-9-91-01 treatment in both genotypes. (b) Endogenous SIK2 was immunoprecipitated from 1 mg liver extracts of control (LKB1lox/lox) or liver-specific LKB1 knockout (LKB1 KO) mice using SIK2 antibody or pre-immune immunoglobulin G (IgG) and subjected to either in vitro kinase assay or immunoblot analysis. Pre-immune IgG was used as negative control. Data is presented as mean±s.d., n=3. Same liver extracts (40μg) were included in immunoblot analysis as input. (c) Glucose production from primary hepatocytes of control (LKB1lox/lox) or liver-specific LKB1 knockout (LKB1 KO) mice was measured following 4 μM HG-9-91-01 or 100 μM Bt2-cAMP treatment for 8 h. Lysates from the same experiment were immunoblotted with the indicated antibodies. Data is presented as mean±s.d., n=3. *P<0.01 basal versus each treatment in control and basal versus Bt2-cAMP in LKB1 KO, #P<0.01 basal control versus basal LKB1 KO, Bt2-cAMP control versus Bt2-cAMP LKB1 KO.
