Fig. 5: Construction of Act-ER FOXO3-expressing mice.
From: FOXO3 is a latent tumor suppressor for FOXO3-positive and cytoplasmic-type gastric cancer cells
a Schematic drawing of the strategy for generating Act-ER FOXO3-expressing mice by homologous recombination. The cDNA of Act-ER FOXO3 linked with 2A peptide and EGFP cDNA was introduced into exon 2 of endogenous Foxo3 by homologous recombination. The short arm (SA) and long arm (LA) of the targeting vector. Black triangles indicate loxP. Asterisks indicate mutations at AKT recognition sites. b Immunoblotting of Act-ER FOXO3 and endogenous FOXO3 in cytosol and nuclear fractions of wild-type and Foxo3Act/Act mouse gastric epithelial cells treated with tamoxifen (Tam) are shown. Lamin A/C and tubulin were used as internal controls of nuclear and cytosol proteins, respectively. The immunoblotting experiments were repeated three times with similar results. c The percent survival of Foxo3+/+, Foxo3+/Act, and Foxo3Act/Act mice after treatment with Tam. Arrows indicate the timing of Tam treatment. (n = 10, 10, and 16 for Foxo3+/+, Foxo3+/Act, and Foxo3Act/Act mice, respectively). d Representative photographs of kidney histology sections (H&E) (low-powered magnification) (left) and enlarged images of the boxed area (H&E) and serial sections for FOXO3 immunohistochemistry (IHC) and Apop Tag staining (right) are shown. Wild-type (top) and Tam-treated Foxo3Act/Act mice (bottom). Insets indicate enlarged images. Bars, 1 mm (left) and 100 μm (right). The images are representative of n = 4 biologically independent animals for each genotype. e The blood urea nitrogen (BUN) levels (top) and serum creatinine (CRE) levels (bottom) of wild-type and Foxo3Act/Act mice are shown in bar graphs (mean ± s.d.). Individual data are shown with dots.
