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. 2016 Apr 22;291(25):13394–13409. doi: 10.1074/jbc.M115.693911

FIGURE 2.

FIGURE 2.

24-h aldosterone treatment increases SOCE. A, left panel, representative traces of fluorescence variation in Fluo-4/AM-loaded NRVMs incubated in the absence (light gray trace), in the presence of Aldo (100 nm, dark gray trace; 1 μm, red trace), or in presence of 1 μm Aldo + 10 μm RU (the selective MR antagonist; orange trace). Cells were exposed to 2 μm Tg + 10 mm caf in the presence of 10 μm NIF in Ca2+-free medium and then to Ca2+-containing solution to evaluate the SOCE. Right panel, scatter plots with mean ± S.E. illustrating amplitude of SOCE (F/F0) in NRVMs incubated for 24 h in the absence (ctrl; open circles) or presence of Aldo (triangles; 10 nm, light gray; 100 nm, dark gray; 1 μm, red) without or with a 10 μm concentration of the selective MR antagonist RU (gray circles and orange triangles). Error bars represent S.E. ***, p < 0.001 versus ctrl; ###, p < 0.001 versus Aldo; n = 3–15 primary cultures; n = 79–250 investigated cells. B, left panel, representative traces of SOCE in Fluo-4/AM-loaded NRVMs incubated in the absence (light gray trace), presence of 1 μm Aldo (red trace), presence of 1 μm Aldo + 40 μm SKF-96365 (SKF; yellow trace), or presence of 1 μm Aldo + 5 μm BTP2 (light blue trace). Right panel, quantitative assessment of SOCE in the ctrl (open circles) or Aldo condition (triangles) in the absence or presence of SOC inhibitor (5 μm BTP2 or 40 μm SKF-96365). Error bars represent S.E. ***, p < 0.001 versus ctrl; ###, p < 0.001 versus Aldo; n = 3–15 primary cultures; n = 64–250 investigated cells. C, left panel, representative fluorescence traces of the Fura-2 decay phase upon Mn2+ addition in untreated (gray trace) or 1 μm Aldo-treated (red trace) NRVMs. Right panel, the initial slope of the Mn2+-induced decrease of Fura-2 fluorescence was fitted by linear regression. Scatter plots with mean ± S.E. illustrate (ΔF/dt) for NRVMs incubated for 24 h with or without 1 μm Aldo. Error bars represent S.E. ***, p < 0.001 versus ctrl; n = 3 primary cultures; n = 76–95 investigated NRVMs. a.u., arbitrary units.