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. 2019 Mar 1;8(5):e011628. doi: 10.1161/JAHA.118.011628

Figure 4.

Figure 4

Effect of 2ME2 on the cellular proliferation of female hPASMCs. Effect of 2ME2 (100 nm–10 μmol/L) on fetal bovine serum–induced proliferation in female hPASMCs assessed by manual cell counts (A), cell counting kit 8 assay (B) and proliferating cell nuclear antigen protein expression after 48 hours (C) (n=4–5). Effect of preincubation with of the ERα antagonist MPP (100 nmol/L), ERβ antagonist PHTPP (100 nmol/L) and G protein–coupled estrogen receptor 1 antagonist G15 (100 nmol/L)±2ME2 (10 μmol/L) in non‐PAH female hPASMCs (D) (n=4). Data are shown as mean±SEM. *P<0.05; **P<0.01; ***P<0.001 as indicated. Statistical analysis: (A through D) determined by 1‐way ANOVA followed by Tukey's post hoc test; panel C determined by paired t‐test. 1%, 1% serum; 5%, 5% serum. Membranes were cut at an appropriate point and probed for separate antibodies. Membranes were then stripped and reprobed with β‐actin. 2ME2 indicates 2‐methoxyestradiol; ERα, estrogen receptor‐alpha; ERβ, estrogen receptor‐beta; hPASMCs, human pulmonary arterial smooth muscle cells; Veh, vehicle.