Journal: PLoS ONE

Article Title: Genetic Ablation of Afadin Causes Mislocalization and Deformation of Paneth Cells in the Mouse Small Intestinal Epithelium

doi: 10.1371/journal.pone.0110549

Figure Lengend Snippet: ( A ) Western blotting of afadin and EphB3 using immunoprecipitates with the indicated antibodies from the Ls174T colon cancer cell line. ( B ) Immunostaining of the small intestine of control and afadin -cKO mice with antibodies against EphB3 (green), lysozyme (red), and Hoechst33258 (blue). Scale bars = 50 µm. ( C ) Relative intensity of EphB3 signal in crypt cells to that of non-crypt cells is shown. 18 crypts from 6 pictures (control) and 24 crypts from 8 pictures ( afadin -cKO) were analyzed.

Article Snippet: The colon cancer cell line Ls174T (DS Pharma Biomedical Co., Osaka, Japan) was cultured in MEM containing 1% NEAA, 2 mM L-glutamine, and 10% FBS and lysed in 50 mM Tris HCl (pH 7.5), 150 mM NaCl, 1 mM MgCl 2 , 1% Nonidet P-40, 1 mM EGTA, and 10% glycerol supplemented with 1 μg/ml aprotinin, 1 μg/ml leupeptin, 20 μg/ml phenylmethylsulfonyl fluoride, and phosphatase inhibitors.

Techniques: Western Blot, Immunostaining, Control