(A) mLEC and MDCK cells were cultured on plastic dishes for several days. After washing with PBS, cells were processed for SDS–PAGE and western blotting as described in METHOD section. Anti-E-Cadherin antibody (cat #3195) was purchased from Cell Signaling. Arrowhead denotes the band corresponding to E-Cadherin. (B) and (C) mLEC and MDCK cells were cultured on a glass-bottom dish for 48 h, washed with PBS, and fixed with 4% paraformaldehyde. Cells were treated with 0.1% Triton X-100 /PBS for 10 min, and treated with Blocking buffer (Nacalai). Cells were incubated with E-Cadherin antibody, followed by the secondary antibody conjugated with Alexa488 (green), Hoechest 33,342 (blue), and phalloidin-conjugated with Alexa 594. Fluorescent images were obtained with Olympus IX-80. Images in (B) and (C) were obtained with x10 (lower magnification), and x60 lenses, respectively. In mLEC cells, E-Cadherin fluorescent intensity was less compared to MDCK cells, and signals come from the nucleus (B and C). In a negative control (without E-cadherin antibody, but with secondary antibody), such fluorescent signal were not observed. To access the data, click or select the words “Appendix 5”