AQP2 expression is not changed in short-term lithium-treated PKCα KO mice.

<p>A) IM tissue collected from injected WT and PKCα KO mice was subjected to Western blot analysis and probed for AQP2. Representative blots showing both nonglycosylated (29-kDa) and glycosylated (35- to 50-kDa) AQP2 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0101753#pone.0101753-Terris1" target="_blank">[59]</a>, and the corresponding loading control, β tubulin. Each lane represents one animal. B) Combined densitometry of all forms of AQP2 in the IM normalized to β tubulin. C) OM tissue probed for AQP2 and β tubulin. D) Combined densitometry of all forms of AQP2 in the OM normalized to β tubulin. Data are presented as percent difference in expression from Day 0, untreated mice as mean ± SEM where *  =  p<0.05 vs. WT day 0 and §  =  p<0.05 vs. PKCα KO day 0 is deemed significant. <i>n = 6</i>.</p

Lithium-induced NDI is attenuated in PKCα KO mice.

<p>WT and PKCα KO mice were provided standard chow or chow containing lithium (40 mmol/kg) for 6 weeks. Single animals were subsequently placed in individual metabolic cages to determine 24-h water intake. Urine and serum were also collected at this time point for metabolic determinations. Uprot/Uosm  =  urinary protein/urine osmolality ratio, CrCl/BW  =  creatinine clearance normalized to body weight (29–30 g). Data are presented as mean ± SEM where *  =  p<0.05 vs. WT day 0 and §  =  p<0.05 vs. PKCα KO day 0 is deemed significantly different. <i>n = 5</i>.</p

UT-A1 expression is not changed in short-term lithium-treated PKCα KO mice.

<p>IM tissue collected from injected WT and PKCα KO mice was subjected to Western blot analysis and probed for UT-A1. A) Representative blots showing the multiple glycosylated forms of UT-A1 that span between 97 and 117 kDa <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0101753#pone.0101753-Chen1" target="_blank">[60]</a>, and the corresponding loading control, β tubulin. Each lane represents one animal. B) Combined densitometry of all glycosylated forms of UT-A1 normalized to β tubulin. Data are presented as percent difference in expression from Day 0, untreated mice as mean ± SEM where *  =  p<0.05 vs. WT day 0 and §  =  p<0.05 vs. PKCα KO day 0 is deemed significant. <i>n = 6</i>.</p

UT-A1 expression is not altered in long-term lithium treatment PKCα KO mice.

<p>IM tissue collected from lithium-fed WT and PKCα KO mice was subjected to Western blot analysis and probed for UT-A1. A) Representative blots showing the multiple glycosylated forms of UT-A1 (bracketed) and corresponding loading control, β tubulin. Each lane represents one animal. B) Combined densitometry of all glycosylated forms of UT-A1 normalized to β tubulin. Data are presented as mean ± SEM where *  =  p<0.05 vs. WT day 0 and §  =  p<0.05 vs. PKCα KO day 0 is deemed significant. <i>n = 12</i>.</p

PKCα KO mice are resistant to lithium-induced natriuresis, kaliuresis and hypercalciuria.

<p>Urinary sodium (A), chloride (B), potassium (C) and calcium (D) were measured and normalized to urinary creatinine to examine difference between lithium-fed WT and PKCα KO to untreated control groups. Data are presented as mean ± SEM where *  =  p<0.05 untreated vs. 6 week-fed lithium treatment. <i>n = 12.</i></p

PKCα KO mice do not develop polyuria in response to short-term lithium treatment.

<p>PKCα KO mice and littermate controls (WT) were injected daily with 40 mmol/kg of lithium for 3 or 5 consecutive days. Urine was collected via metabolic cages and urine osmolality was determined. Data are presented as mean ± SEM where *  =  p<0.05 vs. WT day 0 and §  =  p<0.05 vs. PKCα KO day 0 is deemed significant. <i>n = 6</i>.</p

Acute lithium treatment does not induce NDI in PKCα KO mice.

<p>PKCα KO and WT mice were injected intraperitoneally with 40 mmol/kg LiCl in saline every 24 hours up to 3 or 5 days. Single animals were subsequently placed in individual metabolic cages and urine and serum were collected after 0, 3 or 5 days of daily lithium treatments. Uprot/Uosm  =  urinary protein/urine osmolality ratio, CrCl/BW  =  creatinine clearance normalized to body weight (23–25 g). Data are presented as mean ± SEM where *  =  p<0.05 vs. WT day 0 and §  =  p<0.05 vs. PKCα KO day 0 is deemed significantly different. <i>n</i> = 6.</p

Long-term lithium treatment does not lower AQP2 expression as extensively in PKCα KO mice.

<p>IM and OM tissues collected from lithium-fed WT and PKCα KO mice were subjected to Western blot analysis and probed for AQP2. A) Representative blots showing both nonglycosylated and glycosylated AQP2 (bracketed) in the IM and the corresponding loading control, β tubulin. Each lane represents one animal. B) Combined densitometry of all forms of AQP2 in the IM normalized to β tubulin. C) A representative blot where each lane represents OM tissue from one animal probed for AQP2 (bracketed) and β tubulin. D) Combined densitometry of all forms of AQP2 in the OM normalized to β tubulin. Data are presented as mean ± SEM where *  =  p<0.05 vs. WT day 0 and §  =  p<0.05 vs. PKCα KO day 0 is deemed significant. <i>n = 12</i>.</p