Figure 5 : Long-term insulin treatment stimulates CRY1 expression and inhibits hepatic gluconeogenesis.

From: SREBP1c-CRY1 signalling represses hepatic glucose production by promoting FOXO1 degradation during refeeding

Figure 5

(a,b) Mouse primary hepatocytes were treated with 10 nM insulin for different periods. Protein levels (a) were determined with western blotting. The mRNA levels (b) were analysed by qRT-PCR. Data are represented as mean ±s.d., N=3 for each group. *P<0.05, ***P<0.001 (Student’s t-test). (ce) Mouse primary hepatocytes isolated from CRY1+/+ and CRY1−/− mice were treated with 10 nM insulin for different periods. Protein levels (c) were analysed by western blotting, and relative mRNA levels (d) were determined by qRT-PCR and normalized to the TBP mRNA level. Data are represented as mean ±s.d., N=3 for each group. *P<0.05, **P<0.01, ***P<0.001 versus CRY1+/+ control (Student’s t-test). Relative glucose production (e) was measured using a glucose oxidase (GO) kit. Data are represented as mean ±s.d., N=5 for each group. *P<0.05, **P<0.01, versus CRY1+/+ control (Student’s t-test). (f) Mouse primary hepatocytes were infected with Ad-MOCK and Ad-CRY1, and then treated with insulin (10 nM) or insulin (10 nM) and AKTVIII (5 μM) for 12 h. Protein levels were determined with western blotting. (gi) C57BL/6 mice were infected with Ad-MOCK or Ad-CRY1 and subjected to the pyruvate tolerance test (g) with or without the AKT inhibitor MK2206. MK2206 (30 mg kg−1) was given by oral gavage 10 min before the pyruvate tolerance test. All mice were fasted at ZT 10 and performed PTT at ZT 3. Results were converted to AUC values (h). After the pyruvate tolerance test, hepatic protein levels (i) were analysed by western blotting. Data are represented as mean ±s.d., N=5–7 for each group. ##P<0.01, ###P<0.001 versus Ad-MOCK+vehicle control, *P<0.05, **P<0.01, versus Ad-MOCK+MK2206 control (Student’s t-test). See Supplementary Fig. 13 for original full immunoblot.