Dysregulation of insulin receptor substrate 2 in β cells and brain causes obesity and diabetes
J. Clin. Invest. Xueying Lin, et al. 114:908 doi:10.1172/JCI22217 [
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Figure 1Creation of
fIrs2 mice and
cr2-mediated recombination/deletion of
fIrs2. (
A) A schematic diagram of the 4 steps in the generation of the
fIrs2 containing embryonic stem cells (see Methods for details). Homol recomb, homologous recombination. (
B) Recombination/deletion of
fIrs2 was revealed in various mouse tissues at 4 weeks of age by PCR analysis of genomic DNA. L, liver; K, kidney; H, heart; S, spleen; B, brain, F, fat; M, muscle. The intact
fIrs2 allele (
fIrs2+) yields a 750-bp fragment, whereas the recombined/deleted allele (
fIrs2_) yields a 250-bp fragment. (
C) Real-time PCR analysis of
Irs2 expression in isolated islets, normalized by
cyclophilin expression. (
D) PCR analysis of hypothalamic DNA to reveal recombination event. (
E) Immunoprecipitation and Western blot analysis of hypothalamus with antibodies against Irs2. (
F) Brain sections were immunostained with antibodies against Irs2 (red) and Cre (green), and the images were merged in Openlab Image software (version 3.14; Improvision). Similar results were obtained with multiple sections from 2 to 8 mice. Asterisks indicate third ventricle. Magnification, ×20.
