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. 2021 Jun;191(6):1135–1150. doi: 10.1016/j.ajpath.2021.03.009

Figure 6.

Figure 6

Relative expression of Elk1 in the lung of female and male EHITSN-K0ITSN+/– mice and human PAECPAH. A–C: Real-time quantitative PCR of mRNA isolated from female and male wild-type (wt) and EHITSN-K0ITSN+/– mouse lungs (A), 5 mice per experimental condition, 5 failed donors (FDs), and 10 human PAH lungs (B), and wt and Sugen5416 inhibitor/hypoxia (Su/Hx) rats (C), 3 rats per experimental condition. Data are representative of three independent experiments performed in triplicate. D: Western blot (WB) analyses of Elk1 protein expression in the lung lysates of female and male wt and EHITSN-K0ITSN+/– mice, three randomly selected PAEC lines of PAH patients and FD subjects, and wt and Su/Hx rats, as indicated. Identical detection conditions were used for WB analyses and electrochemiluminescence detection. Densitometry shows the average of Elk1/actin ratios in controls and PAH conditions. P value shown for EHITSN-K0ITSN+/– females and males versus the corresponding wt mice, and for PAH human female PAECs versus female ND-Ctrl cells; no significant differences were recorded for human PAH male PAECs versus ND-Ctrl male cells and for Su/Hx male and female rats versus the corresponding wt controls. Elk1 and actin immunoreactivities detected in the control samples and different experimental conditions (EHITSN, human PAECPAH, and Su/Hx rat), as indicated, are separated by vertical lines. Human PAEC lines (from Pulmonary Hypertension Breakthrough Initiative): idiopathic PAH: 5 White females (average age, 30.6 years; range, 16 to 47 years); 5 males (3 White, 2 Black) (average age, 48.8 years; range, 27 to 54 years); FDs: 3 White females (aged 36, 49, and 55 years); 2 White males and 1 of unknown race (aged 25, 20, and 30 years); all non-Hispanic. Data represent means ± SD (A–C). n = 3 independent experiments (D). ∗P < 0.05. Elk1, ETS domain containing protein; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; ND-Ctrl, nondisease controls; PAECPAH, pulmonary artery endothelial cells of pulmonary arterial hypertension patients.