FIGURE 5.

Depletion of cholesterol reduces the strength of well established cell-cell junctions. Dispase-based dissociation assay revealed that a diminished concentration of membrane cholesterol loosened cell-cell junctions and, consequently, reduced the number of cell-cell junctions. Each column represents the ratio between the number of dissociated cells in the MCD medium and the number of separated cells in the control medium. The proportion of dissociated cells increases in correlation with the duration of the MCD treatment (0.5 h < 1 h < 4 h). Images above the columns represent the corresponding Dsc2 distribution. Thirty minute (A) and 1-h (B) incubation in MCD showed a continuous Dsc2 distribution, whereas after a 4-h incubation (C, arrows) in MCD, a discontinuous Dsc2 distribution was found at places at which the cells had become disconnected. The strength of cell-cell junctions was restored when MCD loaded with cholesterol (MCD-CHOL) was applied to cells for 1 h (D). Error bars represent the S.E. from an experiment in which each condition was tested three times in triplicate.