Fig. 5.
Downregulated-in-adenoma (DRA) expression in the stomach. A: total tissue lysate was prepared from the nonglandular and glandular regions of the stomach and used for Western blot detection. Protein (30 μg) was separated on 8% SDS-PAGE and immunoblotted with DRA antibody (1:1,000 dilution) and GADPH antibody (1:10,000 dilution). B: stomach tissue sections were prepared and reacted with DRA antiserum at 1:100 dilution. Immunohistochemical staining results were analyzed by MRC-1024ES laser-scanning confocal microscopy. C: RNA was isolated from the fundic glands region of the stomach and used for PCR analysis. DRA mRNA and TBP mRNA were amplified with mouse-specific primers. The changes in DRA gene expression were analyzed by the Ct method. Data are means ± SE from a total 10 mice. *P = 0.0002 for NHE8−/− mice vs. wild-type mice. D: total tissue lysate was prepared from the fundic glands region in the stomach and used for Western blot detection. Protein (30 μg) was separated on 8% SDS-PAGE and immunoblotted with DRA antibody (1:1,000 dilution) and β-actin antibody (1:5,000 dilution). The expression of DRA protein was calculated by dividing the optical density of the DRA band over that of the β-actin band. The bar chart shows the DRA protein expression indicated as means ± SE in the sum of a total of 12 mice. *P = 0.0003 for NHE8−/− mice vs. wild-type mice. Inset, the corresponding Western blot image.