Fig. 3. Assessment of junction-associated protein distribution. (A-B) E-cadherin and -catenin levels are not reduced in the Dia1-knockdown cells. (A) Western blot of cell lysates from cells stably expressing the shRNA against LacZ (cont; lane 1) and cells stably expressing the shRNA against hDia1 (lane 2) blotted for E-cadherin and -catenin. Blotting for -tubulin was used as a loading control. (B) Quantitative measurement of Dia1, E-cadherin and -catenin levels, adjusted for the difference in gel loading. Values for control cells were taken as 100%. (C,D) The fluorescence intensity of E-cadherin and actin at cell-cell contacts is reduced in the Dia1-knockdown cells. Representative plot profiles of E-cadherin fluorescence intensity at cell-cell contacts in control (C) and knockdown (D) cells. The white boxes in Fig. 1H,K mark the regions corresponding to plots C and D, respectively. (E) Quantification of the fluorescence intensity of E-cadherin and actin at cell-cell contacts measured by line-scan analysis on digital images. Bars represent the average fluorescence intensity expressed in arbitrary units. Error bars represent s.d. values. Asterisks indicate the values that differ significantly from corresponding controls (Student's t-test, P<0.0001).