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. 2016 May 23;11(5):e0156021. doi: 10.1371/journal.pone.0156021

Fig 5. Spilanthol decreases apical plasma membrane expression of AQP2 in freshly isolated kidney slices and MCD4 cells.

Fig 5

(A) Immunofluorescence analysis of AQP2 subcellular localization in freshly isolated kidney slices in resting condition (Ctr), after 100 μg/ml spilanthol stimulation (Spil), and after incubation with dDAVP in the absence (dDAVP) or in the presence of the spilanthol (Spil + dDAVP). Compared to Ctr and Spil conditions, dDAVP redistributed AQP2 staining to the apical plasma membrane. Of note, spilanthol prevented the dDAVP-induced effect on AQP2 membrane accumulation. Bar = 30 μm. (B) Immunofluorescence analysis of AQP2 subcellular localization in MCD4 renal cells in resting condition and after incubation with forskolin (FK) in the absence or in the presence of 100 μg/ml spilanthol (Spil + FK). AQP2 immunostaining was visualized in the xy apical confocal plan (upper panels) and in the xz confocal plan (lower panels). Compared to Ctr conditions, FK redistributed AQP2 staining to the apical plasma membrane. Spilanthol prevented the FK-induced effect on AQP2 membrane accumulation. Bar = 20 μm. Pictures are representative of at least three independent experiments giving the same results.