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. 2019 Jun 6;4(11):e126910. doi: 10.1172/jci.insight.126910

Figure 4. SGK1 activity is required for K+ stimulation of ENaC.

Figure 4

(A and B) K+ stimulates SGK1 phosphorylation in mpkCCD cells. Cells from Figure 3, B and C, were lysed and prepared for Western blot, and stained with antibodies as indicated. Upper panels: Western blot images showing blots stained with anti–phospho-SGK1 S422, total SGK1, and α-tubulin as labeled. Lower panels: Quantitation of phospho-SGK1/total SGK1 (as described in Methods). Data are mean ± SEM from 3 independent experiments. *P < 0.05, ***P < 0.001 by 1-way ANOVA (A, lower panel). NS, not significant. *P < 0.05 by 2-tailed Student’s t test (B, lower panel). (C and D) Inhibition of SGK1 activity decreases K+-induced ENaC activity. mpkCCD cells were adapted to 1 mM (C) or 3 mM (D) extracellular [K+] as in Figure 3, B and C. The SGK1 inhibitor GSK650394 or vehicle was added and extracellular [K+] was then increased by addition of KCl for 1 hour prior to determining amiloride-sensitive Na+ current. Data are mean ± SEM from 3 independent experiments. ***P < 0.001, ****P < 0.0001 by 1-way ANOVA. μA, microamperes.