Central insulin action regulates peripheral glucose and fat metabolism in mice
J. Clin. Invest. Linda Koch, et al. 118:2132 doi:10.1172/JCI31073 [Go to this article.]

Figure 3
Generation of IR^Δwb mice. (A) General scheme of the inducible whole body IR knockout mouse strain. Expression of an IR-specific shRNA is dependent on the RNA-polymerase III–dependent (Pol III–dependent) promoters H1/U6 containing the operator sequences (tetOs) of the E. coli tetracycline resistance operon. Binding of the tetracycline repressor (tetR) to tetO prevents transcription. Doxycycline (Dox) sequesters tetR and enables the binding of polymerase III to the H1/U6 promoter, which results in transcription of the shRNA. Binding of the shRNA to complementary mRNAs results in the degradation of the IR mRNA. (B) Western blot analysis of IR and AKT (loading control) in whole brain, skeletal muscle, liver, and WAT of 11- to 15-week-old Control^Δwb, IR^Δwb, Control^Δper, and IR^Δper mice. Tissues were dissected 30 days after the start of inducer administration, except for WAT of Control^Δwb and IR^Δwb mice, which was extracted 7 days after start of doxycycline administration. Animal groups were comprised of 3-4 IR^Δwb, 3–4 Control^Δwb, 7–15 IR^Δper, and 9–17 Control^Δper mice. (C) Comparative densitometric analysis of IR expression of 11- to 15-week-old Control^Δper (black bars), IR^Δper mice (white bars), Control^Δwb (dark gray bars), and IR^Δwb mice (light gray bars) of the western blot analysis shown in B. Values are mean ± SEM. *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001 versus control. ANOVA values: brain, 0.027; muscle, 0.004; liver, 0.000; WAT, 0.000.