Generation of pGrb10KO mice. A: Schematic diagram of the strategy used to target the mouse Grb10 locus. The targeting vector is shown. B: Southern blot analysis of WT, homozygous (Ho), and heterozygous (Het) Grb10 floxed mice. Genomic DNA extracted from the
mice was digested with EcoRV and gel-fractionated. The blot was hybridized with 5′- or 3′-probe as indicated. C: Western blot analysis of tissues from pGrb10KO and control mice to confirm the successful knockout of Grb10 in the pancreas.
ERK1/2 is shown as a loading control. D: The pancreases of pGrb10KO (KO) and WT littermates were sectioned at 10 µm by using a cryostat. Protein expression in pancreas
sections was determined using goat anti-insulin (C-12) antibody (Santa Cruz Biotechnology) and a homemade, affinity-purified
rabbit polyclonal antibody to Grb10 (8). The slide images were obtained using a laser confocal fluorescence microscope. Scale bars, 40 µm. B, brain; F, fat; H,
heart; K, kidney; L, liver; Lu, lung; M, muscle; P, pancreas; S, spleen. (A high-quality digital representation of this figure
is available in the online issue.)