Fig 2: ET-1-mediated vasoconstriction, contribution of ETB to ET-1-mediated response and coronary tissue levels of ET-1 and ETB in 4-month-old male and female offspring. (i) ET-1-mediated vasoconstriction in the presence or absence of (a) BQ123 (a competitive antagonist of ETA) or (b) BQ788 (a selective ETB receptors antagonist) in left descending coronary arteries of Normoxia (red) and p-Hypoxia (blue) male (A; circles) and female (B; squares) 4-month-old offspring. (ii) Data are summarized as area under the curve (AUC); n = 4–9 dams/group. (c) Representative confocal images of (i) ET-1 (green), (ii) ETA (green) and (iii) ETB (green) co-stained with CD31 (endothelial cell marker; red) and quantitative analysis of immunofluorescence staining in Normoxia (red symbol) and p-Hypoxia (blue symbol) male (circles) and female (squares) offspring; n = 4–6 dams/group. Data are presented as mean ± SEM; analyzed by t-test or two-way ANOVA with Sidak’s multiple comparison post-hoc test, ** p < 0.01, **** p < 0.0001.

Fig 3: Immunohistochemistry and mRNA expression of AGTR1, ET-1, and eNOS in the thoracic aorta. (A) protein expressions of AGTR1, (B) protein expressions of ET-1, (C) protein expressions of eNOS, (D) mRNA expression of AGTR1, (E) mRNA expression of ET-1, and (F) mRNA expression of eNOS. (× 200 Magnification, scale bar: 100 µm). The results are reported as means ± standard deviations. ***P < 0.001 versus WKY, #P < 0.05, ##P < 0.01, ###P < 0.001 versus SHR, and ?P < 0.05 versus CAP groups. (n = 8).

Fig 4: Immunohistochemistry and mRNA expression of AGTR1, ET-1, and eNOS in the kidney. (A) protein expressions of AGTR1, (B) protein expressions of ET-1, (C) protein expressions of eNOS, (D) mRNA expression of AGTR1, (E) mRNA expression of ET-1, and (F) mRNA expression of eNOS. (× 200 Magnification, scale bar: 100 µm). The results are reported as means ± standard deviations. ***P < 0.001 versus WKY, #P < 0.05, ##P < 0.01, ###P < 0.001 versus SHR, and ?P < 0.05 versus CAP groups. (n = 8).

Fig 5: ET-1-mediated vasoconstriction, contribution of ETB to ET-1-mediated response and coronary tissue levels of ET-1 and ETB in 9.5-month-old male and female offspring. ET-1-mediated vasoconstriction in the presence or absence of (a) BQ123 (a competitive antagonist of ETA) or (b) BQ788 (a selective ETB receptors antagonist) in left descending coronary arteries of Normoxia (red) and p-Hypoxia (blue) male (A; circles) and female (B; squares) 9.5-month-old offspring. (ii) Data are summarized as area under the curve (AUC); n = 5–8 dams/group. (c) Representative confocal images of (i) ET-1 (green), (ii) ETA (green), and (iii) ETB (green) co-stained with CD31 (endothelial cell marker; red) and quantitative analysis of immunofluorescence staining in Normoxia (red symbol) and p-Hypoxia (blue symbol) male (circles) and female (squares) offspring; n = 5–6 dams/group. Data are presented as mean ± SEM; analyzed by t-test or by two-way ANOVA with Sidak’s multiple comparison post-hoc test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.