Fig. 3

Involvement of hPIP in hAQP5 trafficking. (A-B) Cells were pretreated for 24 h with 50µM indomethacin prior to treatment for 8 h without (CTRL) or with 10 µM FK, 0.1 µM TH, or both (FK + TH), in 3 independent experiments. (A) Violin plots (with median and interquartile range of 25–75% percentile) of the membrane upper quartile (UpQ) intensity. Statistical significance evaluated using Kruskal-Wallis test with post-hoc Dunn’s tests is indicated as follow: *: p < 0.05, **: p < 0.01, ****: p < 0.0001. In the absence of PIP, the medians with IQR are: 1 with IQR 0.709–1.287, n = 482 cells for CTRL; 1.274 with 0.872–1.817, n = 721 cells for FK; 1.787 with IQR 1.367–2.192, n = 305 cells for TH; 1.493 with IQR 0.890–1.986, n = 651 cells for FK + TH. In the presence of PIP, the medians with IQR are: 1.553 with IQR 1.075–2.005, n = 606 cells for CTRL; 1.335 with IQR 0.722–2.196, n = 596 cells for FK; 1.651 with IQR 0.875–2.216, n = 565 cells for TH and 1.717 with IQR 1.136–2.344, n = 594 cells for FK + TH. Schematic representations of SNAP-hAQP5 interaction with hPIP was created with BioRender.com. (B) Representative confocal images of immunofluorescent staining with anti-AQP5 antibodies (red) and DAPI (blue); yellow arrows indicate AQP5 localization close to the plasma membrane