Journal: Frontiers in Physiology
Article Title: The E3 ubiquitin-protein ligase Nedd4-2 regulates the sodium chloride cotransporter NCC but is not required for a potassium-induced reduction of NCC expression
Figure Lengend Snippet: Kir4.1, Kir5.1, and pS373-SPAK levels are increased in tubule suspensions isolated from Nedd4-2 KO mice. (A+B) Semi-quantitative assessment of Kir4.1, Kir5.1, and pS373-SPAK levels in tubules isolated from control and Nedd4-2 KO mice treated with 0.5 mM, 3.5 mM, or 8.0 mM K + for 24 h. (A) Data are means ± S.E.M and normalized to control tubules treated with 3.5 mM K + . Difference between control and KO tubules treated with either 0.5 mM, 3.5 mM, or 8.0 mM K + is assessed by 2way ANOVA followed by Tukey’s multiple comparisons test (* p
Article Snippet: Immunoblotting was performed using standard methods and the following primary antibodies: rabbit polyclonal antibodies against NCC (SPC-402D, StressMarq), phosphorylated Threonine-58 NCC (pT58-NCC) , Nedd4-2 , phosphorylated Serine373 SPAK/Serine325 OSR1 (pS373-SPAK) (07–2273, Sigma), SPAK/OSR1 , Kir4.1 (APC-035, Alomone), Kir5.1 (LS-C177333, LS Bio), Proteasome 20S (ab3325, Abcam) and Actin (A2066, Sigma) plus a mouse monoclonal NCC antibody ( ) .
Techniques: Isolation, Mouse Assay