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. 2012 Sep;82(3):500–509. doi: 10.1124/mol.112.079293

Fig. 3.

Fig. 3.

RGS2 protein was stabilized by ouabain and digoxin. HEK-293 cells stably transfected with RGS2-PL were treated overnight with 100 nM ouabain, 100 nM digoxin, or 10 μM MG-132. After the treatment, cycloheximide (10 μg/ml) was added to block protein translation at different time points, to determine the RGS2 protein half-life. A, chemiluminescence corresponding to RGS2 protein levels was detected with the PathHunter ProLabel assay. B, quantification of RGS2 protein half-life (n = 3). The half-life of RGS2 under control conditions was 17.5 ± 5.8 min. Ouabain and digoxin increased the half-life of RGS2 to 77.9 ± 43.4 and 54.4 ± 26.1 min, respectively. Treatment with MG-132 completely stabilized RGS2 expression. *, p < 0.05; **, p < 0.01, using one-way analysis of variance with Bonferroni's post hoc test for multiple comparisons.