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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. 5. Effect of Nedd4-2 on hOAT4 ubiquitination and transport activity.

Fig. 5

(a) hOAT4-expressing cells were transfected with or without wild type Nedd4-2. Transfected cells were then lysed, and hOAT4 was immunoprecipitated (IP) with anti-myc antibody (myc was tagged to hOAT4), followed by immunoblotting (IB) with anti-ubiquitin antibody (anti-Ub). (b) The same immunoblot from Fig. 5a was reprobed by anti-myc antibody. (c) Densitometry plot of results from Fig. 5a, as well as from other experiments. The values are mean ± S.E. (n = 3). *P<0.05. (d) COS-7 cells were co-transfected with hOAT4 and control vector or with hOAT4 and wild type Nedd4-2. 3-min uptake of [3H]-estrone sulfate (0.1 μM) was then measured in these cells. The data represent uptake into hOAT4-transfected cells minus uptake into mock cells (parental COS-7 cells). Values are mean ± S.E. (n = 3). *P<0.05