Fig. 2. Primary Timp3^–/– MECs are more resistant to EGTA-induced cell detachment and have higher -catenin levels. (A) Representative images from at least three different experiments. An increased number of Timp3^–/– epithelial islands (outlined with dotted circles) remain following EGTA treatment as assessed by Hoechst staining and fluorescence microscopy. Bars, 2 mm. (B) Representative western blots of whole-cell lysates from WT and Timp3^–/– (t3^–/–) MECs probed with the indicated antibodies. Quantification by densitometry of (C) E-cadherin signal and (D) -catenin signal (, WT; , Timp3^–/–; ^*P<0.05). (E) Western blot analyses of nuclear lysates of WT and Timp3^–/– MECs using an anti--catenin antibody. (F) MEC total and nuclear lysates subjected to western blotting using an anti-dephosphorylated -catenin (de-P--cat). Panels B-F represent one of three pools of primary mammary epithelial cultures. Each MEC pool was harvested from all ten mammary glands of five WT and five Timp3^–/– female mice. Amido Black and silver staining were used to assess protein loading. Student's t-test was performed to assess statistical significance between WT and Timp3^–/–. Bar graphs were expressed as the mean ± s.e.m.