Fig. 5: Cellular junctions strengthen with RPE polarization.

A Representative 3D reconstructions of an iRPE cell expressing GFP-tagged TJP1 (tight junctions), CTNNB1 (adherens junctions), DSP (desmosomes), and GJA1 (gap junctions). Images from W1 constitute the baseline, while images at W4 show the endpoint for cells that were grown in the presence of PGE2 or HPI4 (magenta = phalloidin, cyan = Hoechst, yellow = GFP-tagged protein; scale bar = 5 μm). Note that the signal from phalloidin was stripped from the top of the cells in these images to better show the reconstruction of the GFP-tagged protein. B Integrated reference maps showing the location of the structure of interest within the cell. The colored points correspond to the average location of each structure of interest from each field of view (n = 30). Each group of points is surrounded by an ellipse, which represents the 90% confidence interval. Cells and nuclei dimensions from Fig. 2 are used as a reference. C, D Raincloud plots indicating the changes of DSP and GJA1 over time and between treatments for C volume and D count per cell (the points indicate the average values for each FOV; the diamonds represent the median, and the error bars indicate the 5th and 95th percentiles. Two-sided Welch’s t-test was performed to compare PGE2- and HPI4-treated cells at each timepoint. N = 30; ***P < 0.005, **P < 0.01, *P < 0.05). The solid lines indicate the generalized linear model maximum likelihood fit for the two treatments with week and the square of week as dependent variables.