Membrane translocation of hNIS protein by cAMP after inhibition of de novo protein synthesis.

Abstract

<p>To inhibit cAMP-induced protein synthesis, AMD (5 ng/mL) or CHX (1 μg/ml) were pretreated 24h before treatment of 100 μM cAMP. (A) Enhanced expression of hNIS/tdTomato proteins by cAMP was visualized after blocking de novo protein synthesis. (B) Enhanced membrane localization of hNIS/tdTomato proteins by cAMP was visualized after blocking de novo protein synthesis. Red fluorescent intensity was analyzed with MetaMorph software. An arbitrary threshold that represented the cytosolic compartment was designated. Threshold intensity of fluorescence was adjusted to show membrane-localized hNIS/tdTomato protein only. Signals over or under the threshold were depicted as red or gray, respectively. (C) The upper threshold of red fluorescent intensity was measured to quantify the membrane localized hNIS/tdTomato protein. Confocal images were collected from at least three different regions of each sample. Bars represent mean ± SD (*, P<0.05; **, P<0.01; N = 3).</p