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Fig. 4. Characterization of gp135 redistribution. (A) Pseudocolor pH maps of two-cell aggregates with compact structure (left panel) and a lumen (middle panel) beside their corresponding fluorescent images used to generate the pH maps. The calibration bar (upper left corner) reports the relative pH, which was normalized to the value measured at the basal surface. Dotted white lines encircle regions corresponding to a compact structure (left panel) or a lumen (central panel). White arrowheads indicate some central vesicles (left panel). Scale bars: 5 µm. The boxplot (right panel) reports the average and relative pH values (small square) ± s.e.m. (box) and the 10-90% quantiles (whiskers) measured in the compact structure, PAP, lumen and central vesicles (n=14). Statistical comparisons were performed on distinct data sets. (B) Intensity profiles of gp135 and caveolin 1 (cav1) or gp58, respectively measured from maximum projections along contact surfaces (x-axis) of two-cell aggregates with PAP and with (upper row) or without (lower row) compact structure. The Gaussian fit of the gp135 signal is reported as a blue curve in the graphs. The corresponding fluorescent channels are reported below the graphs. Bars 5 µm. (C) Frequencies of two-cell aggregates with PAP or with lumen in cultures treated with BFA. Average values of three experiments (± s.e.m., n=50) are shown. BFA caused significant changes in lumen- and PAP-containing aggregates (^*P<0.001).

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