Figure 2.

Conditional AdipoR1 or AdipoR2 gene knockout in β-cells exhibited no effect on glucose metabolism and β-cell proliferation during pregnancy. A: mRNA levels of AdipoR1 and AdipoR2 were measured by real-time PCR with samples from C57BL/6 dams at G18.5. B–M: The AdipoR1^fl/fl or AdipoR2^fl/fl mice were crossed with Ins1^CreERT2 mice to produce female mice with AdipoR1^fl/fl;Ins1^CreERT2 (ibR1ko) or AdipoR2^fl/fl;Ins1^CreERT2 genotype (ibR2ko). Littermates with AdipoR1^−/−;Ins1^CreERT2 or AdipoR2^−/−;Ins1^CreERT2 genotypes were used as control (Con). Tamoxifen was gavaged (1 mg in 100 μL corn oil) from G7.5 to G11.5. B and C: expression levels of AdipoR1 and AdipoR2 were determined by TaqMan PCR with purified islets. D–G: GTT was performed on G12.5 after 6 h fasting (n = 6–8). H and I: β-Cell areas were measured with anti-insulin antibody–probed immunohistochemistry images (image not shown). J–M: Ki67 protein in β-cells were detected by IF with anti-Ki67 antibody (red, indicated with arrowhead) and anti-insulin antibody (green). Nuclei were stained by DAPI (blue); white scale bar, 50 μm. N: Pancreatic islets were isolated from 10-week-old female C57BL/6 mice. After overnight incubation, islets were treated with purified adiponectin (2 μg/mL), bPL (0.1 μg/mL), or BSA for 48 h. The islets were trypsinized, and cells were loaded to coated slides. IF was performed with use of anti-Ki67 and anti-insulin antibody. We calculated the percentages of Ki67-positive β-cells calculated using IF images. Data are presented as mean ± SEM. AU, arbitrary units; S.Muscle, skeletal muscle.