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

Fig 2. Exposure to spilanthol reduced basal NKCC2 phosphorylation (pNKCC2) in mouse kidney slices.

Fig 2

(A) Coimmunostaining of R5 antibody (pNKCC2) with AQP2 as marker of collecting ducts (AQP2) and Tamm-Horsfall Protein as marker of TAL tubules (THP) in serial kidney sections. Asterisks indicate corresponding TAL tubules in the serial sections, Bar = 30 μm. (B) Kidney slices (250 μm) were stimulated 2h with either 100 μg/ml MPE or 100 μg/ml spilanthol, then lysed and total protein extracts analyzed for pNKCC2 expression as shown by this representative Western blot. Right panel. Densitometric analysis showed a significant reduction of pNKCC2 (normalized to total NKCC2) in kidney slices stimulated with either MPE (***p<0.001) or spilanthol (***p<0.001) compared to control condition. Spilanthol-induced reduction of pNKCC2 was significantly larger than that evoked by MPE (**p<0.01). Comparable results were obtained in two different mice preparations and significance calculated by Student’s T-test for unpaired data.