Figure 5.

USP8 knockdown significantly reduced intercellular communication via GJs. A, flow cytometric analysis of unlabeled (top) and calcein AM–labeled (bottom) scramble or shUSP8-infected MEFs. B, unlabeled MEFs were co-cultured with calcein AM–labeled MEFs (1:1) for 3 h to allow the dye transfer between two populations, and then subjected for the flow cytometric analysis. In mixed culture, 50% cells with low fluorescence intensity present unlabeled population (top panel). The relative mean fluorescence intensity (MFI) of unlabeled cells in shUSP8-infected MEFs was calculated and normalized to that in scramble-infected MEFs (bottom panel). Each circle, square, or triangle represents one experiment. The data are shown as mean ± S.D. from four independent experiments. *, p < 0.05; Student's t test.