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. Author manuscript; available in PMC: 2017 Feb 15.
Published in final edited form as: Biochem Pharmacol. 2015 Dec 29;102:120–129. doi: 10.1016/j.bcp.2015.11.024

Fig. 7. Effect of Nedd4-2 ligase activity on sgk2 regulation of hOAT4 cell surface expression.

Fig. 7

(a). COS-7 cells were transfected with hOAT4, CA-sgk2 and control vector, or with hOAT4, CA-sgk2 and Nedd4-2, or with hOAT4, CA-sgk2 and ubiquitin ligase-dead Nedd4-2 mutant (Nedd4-2/C821A). Transfected cells were labeled with biotin. Biotinylated/cell surface proteins were separated with streptavidin beads, followed by immunoblotting (IB) with an anti-myc antibody. (b) COS-7 cells were transfected with hOAT4, CA-sgk2 and control vector, or with hOAT4, CA-sgk2 and Nedd4-2, or with hOAT4, CA-sgk2 and ubiquitin ligase-dead Nedd4-2 mutant (Nedd4-2/C821A). Transfected cells were then lysed, followed by immunoblotting with anti-actin antibody, anti-sgk2 antibody or anti-Nedd4-2 antibody, respectively. (c). Densitometry plot of results from Fig. 7a, as well as from other experiments. The values are mean ± S.E. (n = 3). *P<0.05, NS: statistically not significant.