Figure 1.

Apoptosis of LLC-PK1 cells producing Y353F ezrin in a tubulogenesis assay. (A) Immunodetection of endogenous and overproduced tagged ezrin in stable LLC-PK1 clones. P, E, and F cells were transfected, respectively, with the empty vector, the same vector encoding VSV-G tagged wild-type ezrin, or VSV-G tagged Y353F ezrin. Total cellular lysates were analyzed with either P5D4 mAb (Tag) or an anti-ezrin serum. (B) Cells were cultured for 7 days in presence of HGF in a collagen type I gel. Cultures were examined with Nomarski optics (Left) and for Hoechst 33258 staining (Right). In a collagen type I gel, only apoptotic bodies, as highlighted by pyknotic nuclei, were observed in F cell cultures, whereas P and E cells were able to form tubules in presence of HGF. (Bar = 20 μm.)