Inner medullary expression of AQP2 and p-AQP2.

<p>Values are expressed as means ± SE. AQP2, aquaporin-2;</p><p>p-AQP2, p(Ser256)-aquaporin-2;</p><p>n, number of rats.</p><p>*<i>P <</i> 0.05 vs. Sham</p><p># <i>P <</i> 0.05 vs. HF</p><p>† <i>P <</i> 0.05 vs. L-HF+d</p><p>‡ <i>P <</i> 0.05 vs. L-Sham</p><p>♣ <i>P <</i> 0.05 vs. L-Sham+d. Values are expressed as means ± SE.</p><p>Inner medullary expression of AQP2 and p-AQP2.</p

Inner medullary expression of AQP1.

<p>Values are expressed as means ± SE. AQP1, aquaporin-1;</p><p>n, number of rats. L-Sham+d.</p><p>*<i>P <</i> 0.05 vs. Sham</p><p>¤ <i>P <</i> 0.05 vs. L-HF.</p><p>Inner medullary expression of AQP1.</p

AQP4 abundance.

<p>Semiquantitative immunoblotting of kidney protein prepared from inner medulla. Immunoblot was reacted with anti-AQP4 antibody and reveals a single ~ 34.5 kDa AQP4-band (<i>A-E</i>). Data are presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0116501#pone.0116501.t010" target="_blank">Table 10</a>. (<i>A-B</i>) Densitometric analysis revealed significantly increased AQP4 protein levels of HF, L-HF and L-Sham vs. Sham. The increase in HF, L-HF and L-Sham was comparable. In contrast, AQP4 protein levels in L-HF+d decreased vs. HF and L-HF in <i>C</i>) and vs. L-Sham and L-Sham+d in <i>D</i>). No difference was observed between L-Sham and L-Sham+d, as also presented in <i>E</i>). <i>E</i>) AQP4 protein levels in L-Sham+d and L-Sham were increased compared with Sham. No difference was observed between L-Sham and L-Sham+d. Each column represents the mean ± SE. Solid white, Sham; solid light grey, HF; line pattern, L-Sham; solid dark grey, L-HF; solid black, L-HF+d; dotted pattern, L-Sham+d. *<i>P <</i> 0.05 vs. Sham, #<i>P <</i> 0.05 vs. HF, † <i>P <</i> 0.05 vs. L-HF+d, ‡ <i>P <</i> 0.05 vs. L-Sham.</p

Inner medullary expression of the Gsα subunit.

<p>Values are expressed as means ± SE. Gsα, Gsα subunit;</p><p>n, number of rats.</p><p>*<i>P <</i> 0.05 vs. Sham</p><p># <i>P <</i> 0.05 vs. HF</p><p>† <i>P <</i> 0.05 vs. L-HF+d</p><p>‡ <i>P <</i> 0.05 vs. L-Sham</p><p>♣ <i>P <</i> 0.05 vs. L-Sham+d.</p><p>Inner medullary expression of the Gsα subunit.</p

AQP2 and p-AQP2 abundance.

<p>Semiquantitative immunoblotting of kidney protein prepared from inner medulla. Immunoblot was reacted with anti-AQP2 (<i>A-E</i>) and AQP2 phosphorylated at Ser256 (p-AQP2) (<i>F-J</i>) antibody. Both antibodies reveal specific 29 kDa and 35–50 kDa bands. Data are presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0116501#pone.0116501.t004" target="_blank">Table 4</a>. Densitometric analysis revealed unchanged AQP2 and p-AQP2 protein levels in HF and L-HF vs. Sham 17 days after MI. Neither sodium restriction nor DDAVP infusion increased AQP2 and p-AQP2 abundance in HF as otherwise observed in L-Sham+d (<i>A</i>, <i>B</i> and <i>F</i>, <i>G</i>). Consistently, L-HF+d revealed decreased AQP2 and p-AQP2 abundance to L-HF levels vs. L-Sham and HF (<i>C</i>, <i>D</i> and <i>H</i>, <i>I</i>). Furthermore, AQP2 and p-AQP2 expression was decreased in L-Sham rats compared with Sham, HF, and L-Sham+d (<i>A</i>, <i>B</i>, <i>E</i> and <i>F</i>, <i>G</i>, <i>J</i>, respectively). In contrast, no difference was observed in L-Sham+d was observed vs. Sham (<i>E</i> and <i>J</i>). Each column represents the mean ± SE. Each column represents the mean ± SE. Solid white, Sham; solid light grey, HF; line pattern, L-Sham; solid dark grey, L-HF; solid black, L-HF+d; dotted pattern, L-Sham+d. *<i>P <</i> 0.05 vs. Sham, # <i>P <</i> 0.05 vs. HF, † <i>P <</i> 0.05 vs. L-HF+d, ‡ <i>P <</i> 0.05 vs. L-Sham, ♣ <i>P <</i> 0.05 vs. L-Sham+d.</p

Inner medullary expression of the V2 vasopressin receptor.

<p>Values are expressed as means ± SE. V2R, V2 vasopressin receptor;</p><p>n, number of rats.</p><p>*<i>P <</i> 0.05 vs. Sham</p><p># <i>P <</i> 0.05 vs. HF</p><p>† <i>P <</i> 0.05 vs. L-HF+d.</p><p>Inner medullary expression of the V2 vasopressin receptor.</p

Gsα subunit abundance.

<p>Semiquantitative immunoblotting of kidney protein prepared from inner medulla. Immunoblot was reacted against anti-Gsα subunit of the G-protein revealing a doublet band at 45 and 50 kDa (<i>A-E</i>). Data are presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0116501#pone.0116501.t006" target="_blank">Table 6</a>. <i>A</i>) Densitometric analysis revealed significantly increased Gsα abundance in HF vs. Sham and L-Sham, whereas Gsα expression was comparable between Sham and L-Sham, as also presented in <i>B</i>) and <i>E</i>). <i>B</i>) Gsα was increased in L-HF vs. Sham and L-Sham. <i>C</i>) Gsα abundance was increased in L-HF rats vs. HF and L-HF+d, whereas Gsα expressions were comparable between HF and L-HF+d. <i>D</i>) Gsα abundance was increased in L-Sham+d rats vs. L-Sham rats and L-HF+d. L-Sham and L-HF+d groups were comparable. <i>E</i>) As observed in <i>D</i>), Gsα abundances in L-Sham+d rats were increased vs. Sham and L-Sham. Each column represents the mean ± SE. Solid white, Sham; solid light grey, HF; line pattern, L-Sham; solid dark grey, L-HF; solid black, L-HF+d; dotted pattern, L-Sham+d. *<i>P <</i> 0.05 vs. Sham, # <i>P <</i> 0.05 vs. HF, † <i>P <</i> 0.05 vs. L-HF+d, ‡ <i>P <</i> 0.05 vs. L-Sham, ♣ <i>P <</i> 0.05 vs. L-Sham+d.</p

Inner medullary expression of the (pro)renin receptor.

<p>Values are expressed as means ± SE. (P)RR, (pro)renin receptor;</p><p>n, number of rats.</p><p>*<i>P <</i> 0.05 vs. Sham</p><p># <i>P <</i> 0.05 vs. HF</p><p>† <i>P <</i> 0.05 vs. L-HF+d</p><p>‡ <i>P <</i> 0.05 vs. L-Sham</p><p>♣ <i>P <</i> 0.05 vs. L-Sham+d.</p><p>Inner medullary expression of the (pro)renin receptor.</p

(Pro)renin receptor abundance.

<p>Semiquantitative immunoblotting of kidney protein prepared from inner medulla. Immunoblot was reacted with a specific antibody against anti-(P)RR revealing a single band at ~ 42 kDa (<i>A-E</i>). Data are presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0116501#pone.0116501.t008" target="_blank">Table 8</a>. (<i>A</i> and <i>B</i>) (P)RR expression was increased in L-HF vs. Sham and L-Sham, whereas no difference in (P)RR protein expression between Sham, HF or L-Sham was found. As also presented in <i>A</i>) and <i>E</i>), no difference was observed between Sham and L-Sham rats. <i>C</i>) (P)RR expression increased in L-HF vs. HF and L-HF+d, whereas no difference was observed between HF and L-HF+d. <i>D</i>) Densitometry revealed significantly decreased (P)RR abundance in L-HF+d and L-Sham+d when compared with L-Sham. The decrease in L-HF+d and L-Sham+d was comparable. <i>E</i>) As already shown, densitometry revealed that L-Sham+d decreased (P)RR expression compared with L-Sham, and no difference was found between Sham and L-Sham. Each column represents the mean ± SE. Solid white, Sham; solid light grey, HF; line pattern, L-Sham; solid dark grey, L-HF; solid black, L-HF+d; dotted pattern, L-Sham+d. *<i>P <</i> 0.05 vs. Sham, # <i>P <</i> 0.05 vs. HF, † <i>P <</i> 0.05 vs. L-HF+d, ‡ <i>P <</i> 0.05 vs. L-Sham, ♣ <i>P <</i> 0.05 vs. L-Sham+d.</p

Changes in renal function.

<p>Values are expressed as means ± SE. The plasma values are measured at the last day of experiment whereas urine values and body weights are measured the day before to avoid error due to anesthesia under echocardiographic measurements. n, number of rats;</p><p>BW, median body weight;</p><p>Water intake, water intake;</p><p>UO, urine output; U-Osm, urine osmolality;</p><p>P-Osm, plasma osmolality;</p><p>U/P Osm, urine-to-plasma osmolality ratio;</p><p>P-Urea, plasma urea;</p><p>U-Urea, urine urea;</p><p>T^ecH₂O, electrolyte free water reabsorption;</p><p>FENa, fractional excretion of sodium into urine;</p><p>UNa x UO, rate of urinary sodium excretion;</p><p>UK x UO, rate of urinary potassium excretion;</p><p>P-Na, plasma sodium;</p><p>P-K, plasma potassium;</p><p>P-Cr, plasma creatinine;</p><p>Ccr, creatinine clearance.</p><p>*<i>P <</i> 0.05 vs. Sham</p><p># <i>P <</i> 0.05 vs. HF</p><p>‡ <i>P <</i> 0.05 vs. L-Sham</p><p>¤ <i>P <</i> 0.05 vs. L-HF</p><p>♣ <i>P <</i> 0.05 vs. L-Sham+d.</p><p>Changes in renal function.</p