Human Samples/Patient Characteristics.

<p>Human Samples/Patient Characteristics.</p

<i>PKG is detected in both canine and human LV samples</i>.

<p>(A) PKG is detected in LV tissue lysates in young normal (Normal), old hypertensive canines (Old HTN) with diastolic dysfunction, as well as bovine lung. (B) PKG is detected by immunoblotting of human LV samples (heart failure (n = 6) and controls (n = 8), as well as bovine lung. Upper blot—HF 1-3, Control 1-4 and lower blot—HF 4-6, Control 5-8 (refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0118664#pone.0118664.t001" target="_blank">Table 1</a>).</p

<i>PDE5 is not detected in LV samples from human controls or patients with heart failure</i>.

<p>(A) PDE5 is not detected by immunoblotting of human LV samples (heart failure (n = 6) and controls (n = 8); bovine lung is used as a positive control (rabbit polyclonal anti-PDE5 antibody provided by Joseph A Beavo, [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0118664#pone.0118664.ref026" target="_blank">26</a>]). Upper blot—HF 1–3, Control 1–4 and lower blot—HF 4–6, Control 5-8 (refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0118664#pone.0118664.t001" target="_blank">Table 1</a>). (B) Western blots of 2-D gels with the anti-PDE5 antibody (Cell Signaling) demonstrate that PDE5 is detected in the bovine lung as 2 isoelectric variants, while PDE5 is not detected in cardiac tissue lysates of human LV samples (control (Control 2)) and heart failure (HF1)).</p

<i>PDE5 is not detected in cardiac LV and RV lysates from canines</i>.

<p>PDE5 was not detected in tissue lysates from the LV or RV in young normal (Normal) or old hypertensive canines (Old HTN) with diastolic dysfunction using the rabbit polyclonal anti-PDE5 antibody provided by Joseph A Beavo, [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0118664#pone.0118664.ref026" target="_blank">26</a>]. Bovine lung is used as a positive control.</p

PDE5 is not detected in murine LV.

<p>(A) Western blots of LV samples from the murine demonstrate that PDE5 is detected in samples from the lung used as a positive control. For LV samples from normal (control), heart failure and compensated hypertrophy, there is non-specific binding of the anti-PDE5 antibody from Santa Cruz at a MW near the positive control, while PDE5 is not detected with the anti-PDE5 antibody from Cell Signaling or the rabbit polyclonal anti-PDE5 antibody provided by Joseph A Beavo, [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0118664#pone.0118664.ref026" target="_blank">26</a>]. (B) Western blots of 2-D gels with the anti-PDE5 antibody (Cell Signaling) demonstrate that PDE5 is detected in the lung as 2 isoelectric variants, while PDE5 is not detected in cardiac tissue lysates (normal mice). However in cardiac tissue lysates, the anti-PDE5 antibody from Santa Cruz demonstrates non-specific binding.</p