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. 2021 Mar 15;131(6):e129433. doi: 10.1172/JCI129433

Figure 6. ER-α is necessary and sufficient for upregulating apelin in vitro.

Figure 6

(A) RV apelin and ER-α protein positively correlate in male and female control and SuHx-PH rats. (B) ER-α siRNA knockdown time course in H9c2 cardiomyoblasts (5 nM; 24 hours prior to E2 [100 nM]). (C) Apelin expression in H9c2 cells treated with ER-α agonist BTP-α (100 nM, 24 hours). (D) Apelin expression in RV cardiomyocytes (RVCMs) isolated from male control rats treated with E2 (10 nM, 24 hours) or BTP-α (100 nM, 24 hours) in vitro. (E) Apelin expression in RVCMs isolated from male and female SuHx-PH rats and treated with BTP (100 nM, 24 hours) in vitro. (F) RV apelin mRNA expression in normoxia, SuHx-PH, or SuHx-PH rats treated with E2 (75 μg/kg/day via s.c. pellets), ER-α agonist PPT (850 μg/kg/day; s.c. pellets) or EtOH vehicle (Veh). (G) Apelin expression in RVCMs isolated from male controls, SuHx-PH, or SuHx-PH rats treated with E2 or PPT in vivo. n = 3 independent experiments for B and C. Cells from n = 3–4 rats/group in D, E, and G. n = 6–8/group in F. B–E and G depict representative Western blots with densitometric analyses. Pearson’s R value and P value shown in A. Dashed lines represent 95% CI. *P < 0.05 versus Scr (scrambled control), $P < 0.05 versus E2 in B; *P < 0.05 versus control in C and D; *P < 0.05 versus Normoxia, #P < 0.05 versus SuHx in E–G. P values by ANOVA with Tukey’s post hoc correction in B, D–G and by Student’s t test in C. Error bars represent mean ± SEM; each data point represents 1 experiment or animal.