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Effect of Torasemide on ligand-dependent transactivation activity of MR in H9C2-MR cells.

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posted on 2013-09-09, 03:10 authored by Basile Gravez, Antoine Tarjus, Ruben Jimenez-Canino, Soumaya El Moghrabi, Smail Messaoudi, Diego Alvarez de la Rosa, Frederic Jaisser

A: The MMTV promoter contains Hormone Response Elements (HRE); the promoter sequence was fused to the Luciferase coding sequence. This construct was transfected into H9C2-MR cells. Upon binding of the aldosterone-MR complexes on HRE, luciferase is transcribed and light emission is enhanced, assessing MR transactivation activity. In the presence of an MR antagonist, binding of aldosterone to MR is prevented and luciferase expression is abolished. B: 10−8 M aldosterone (Aldo) increased MR transactivation activity (luciferase activity) which was prevented by a 100-fold excess of the MR antagonist spironolactone (Spiro; 10−6 M) but not by the GR antagonist RU 38486 (RU 38486; 10−6 M). Each antagonist alone has no effect. Mean ± SEM (n = 8). * p<0.05 vs control (Ctrl); # p<0.05 vs aldosterone. C: 10−8 M aldosterone (Aldo) increased MR transactivation activity, which was fully inhibited by the MR antagonist spironolactone (S) at 10-6 M. Torasemide (T) has a slight antagonist effect independent of its concentration. Mean ± SEM (n = 4). *p<0.05 vs control (Ctrl); # p<0.05 vs aldosterone.