Fig. 1: PLPP/CIN-mediated NEDD4-2 dephosphorylation.
a–c Effect of PLPP/CIN deletion on expression and phosphorylation levels of SGK1 and NEDD4-2 in vivo. Total NEDD4-2 protein and pNEDD4-2 S448 phosphorylation levels are higher in PLPP/CIN−/− mice, as compared to WT mice. a Representative western blots of SGK1 and NEDD4-2. b, c Quantification of SGK1 and NEDD4-2 expression, their phosphorylation levels, and the ratio of pNEDD4-2 to total NEDD4-2. Open circles indicate each individual value. Horizontal bars indicate mean value (mean ± S.E.M.; *p < 0.05 vs. WT animal; n = 7, respectively). d Representative co-immunoprecipitation data revealing PLPP/CIN-NEDD4-2 bindings. e–g In vitro assay using recombinant proteins. e, f Representative co-immunoprecipitation and western blot data demonstrating PLPP/CIN-NEDD4-2 bindings, and PLPP/CIN-mediated NEDD4-2 S448 dephosphorylation. g Quantification of NEDD4-2 phosphorylation levels based on western blot data. Open circles indicate each individual value. Horizontal bars indicate mean value (mean ± S.E.M.; *p < 0.05 vs. vehicle; n = 7). h, i Effect of PLPP/CIN deletion on NEDD4-2 ubiquitination in vivo. Co-immunoprecipitation reveals that PLPP/CIN deletion decreases the ubiquitin-NEDD4-2 bindings. h Representative western blots of ubiquitin-NEDD4-2 bindings. i Quantification of NEDD4-2 ubiquitination based on western blot data. Open circles indicate each individual value. Horizontal bars indicate mean value (mean ± S.E.M.; *p < 0.05 vs. WT animal; n = 7)
