Figure 1.

Generation of FoxO1-S253^A/A mice and postprandial hyperglycemia. (A) The genomic locus of FoxO1 in mouse genome. (B) Gene-targeting strategy for generation of FoxO1-S253A mutation via embryonic stem (ES) cell and homologous recombination (HR). (C and D) Southern blot was performed for screening the 5′- and 3′-end HR in FoxO1 genomic loci. DNA sequencing of endogenous FoxO1 loci of negative and positive ES cells. (E) Body weight curve of mice of control wild-type (WT), heterozygous (A/+), and homozygous (A/A) mice at the ages of 4 to 16 weeks. (F) Effect of insulin on FoxO1 protein and phosphorylation (p) in hepatocytes. Primary hepatocytes were isolated from the control and A/A mice and treated with 100 nM insulin for 30 minutes, and 100 µg protein of cell lysates was subjected to Western blot against the antibody of FoxO1, T24, S316, and β-actin. (G and H) Blood glucose levels were measured in WT, A/+, and A/A mice at the ages 4 to 16 weeks during (G) 16 hours fasting and (H) random-fed conditions. *P < 0.05 vs WT and A/+ mice, n = 10 mice per group. M, Marker; Neo, neomycin; PGK-1, phosphoglycerate kinase 1; TK, thymidine kinase; utr, untranslated region.