The Cx50–AQP0 interaction enhances Cx50 intercellular gap junction coupling using parachute dye transfer assay. Six days after infection with recombinant retroviruses containing RCAS(A), AQP0, Cx50, Cx50 plus AQP0, Cx46, Cx46 plus AQP0, Cx50*46L, Cx50*46L plus AQP0, Cx50*43L or Cx50*43L plus AQP0, a time-course analysis of parachute dye transfer assay was performed at different time points after parachute using Dil as a tracer dye and calcein as a transferring dye. (A) Immunofluorescence images show the calcein transfer emanating from pre-loaded cells through gap junctions into neighboring cells. (B) The extent of dye transfer was quantified by measuring the area of calcein-fluorescence-stained cells and comparing this with the area of DiI-stained cells. The data are presented as mean ± s.e.m. (n=3). (*P<0.05 and **P<0.01, compared with Cx50).