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Primer sequences for qPCR.

Journal: Life

Article Title: Duloxetine-Induced Antidiuresis in Rats with Lithium-Induced Nephrogenic Diabetes Insipidus

doi: 10.3390/life14081012

Figure Lengend Snippet: Primer sequences for qPCR.

Article Snippet: After being blocked with 5% skim milk in PBS-T (80 mM Na 2 HPO 4 , 20 mM NaH 2 PO 4 , 100 mM NaCl, 0.1% Tween-20, pH 7.5) for 1 h, the membranes were probed overnight at 4 °C with one of the following primary antibodies: rabbit polyclonal anti-AQP2 (#AQP-002, Alomone Labs, Jerusalem, Israel), rabbit polyclonal anti-pSer256-AQP2 (#ab111346, Abcam, Cambridge, MA, USA), rabbit polyclonal anti-CREB-1 (#06-863, Sigma-Aldrich), mouse monoclonal anti-p-CREB-1 (#sc-81486, Santa Cruz Biotechnology, Dallas, TX, USA), and mouse monoclonal anti-β-actin (#A5441, Sigma-Aldrich).

Techniques:

Effects of duloxetine treatment on aquaporin-2 (AQP2) protein expression in lithium-induced nephrogenic diabetes insipidus. Immunoblots of AQP2 and pS256-AQP2 are shown; each lane was loaded with a protein sample from a different rat kidney ( A ). Densitometric analysis revealed that the protein levels of AQP2 and pS256-AQP2 were significantly reduced by lithium treatment (Li) compared to controls, but were restored by lithium/duloxetine co-treatment (Li + DX). The levels of AQP2 and pS256-AQP2 proteins increased in normal rats treated with duloxetine (DX) compared to the control group ( B ). Immunohistochemistry is shown for AQP2 ( C ) and pS256-AQP2 ( D ) in rat kidneys. Along the cortical collecting duct (CCD), outer medullary collecting duct (OMCD), and inner medullary collecting duct (IMCD), differences in AQP2 labeling among groups were consistent with those in the immunoblot analyses. Each bar in the densitometry results represents the mean ± standard deviation ( B ). *, p < 0.05 vs. control; # , p < 0.05 vs. Li by the post hoc Mann–Whitney U test. Scale bars, 50 μm.

Journal: Life

Article Title: Duloxetine-Induced Antidiuresis in Rats with Lithium-Induced Nephrogenic Diabetes Insipidus

doi: 10.3390/life14081012

Figure Lengend Snippet: Effects of duloxetine treatment on aquaporin-2 (AQP2) protein expression in lithium-induced nephrogenic diabetes insipidus. Immunoblots of AQP2 and pS256-AQP2 are shown; each lane was loaded with a protein sample from a different rat kidney ( A ). Densitometric analysis revealed that the protein levels of AQP2 and pS256-AQP2 were significantly reduced by lithium treatment (Li) compared to controls, but were restored by lithium/duloxetine co-treatment (Li + DX). The levels of AQP2 and pS256-AQP2 proteins increased in normal rats treated with duloxetine (DX) compared to the control group ( B ). Immunohistochemistry is shown for AQP2 ( C ) and pS256-AQP2 ( D ) in rat kidneys. Along the cortical collecting duct (CCD), outer medullary collecting duct (OMCD), and inner medullary collecting duct (IMCD), differences in AQP2 labeling among groups were consistent with those in the immunoblot analyses. Each bar in the densitometry results represents the mean ± standard deviation ( B ). *, p < 0.05 vs. control; # , p < 0.05 vs. Li by the post hoc Mann–Whitney U test. Scale bars, 50 μm.

Article Snippet: After being blocked with 5% skim milk in PBS-T (80 mM Na 2 HPO 4 , 20 mM NaH 2 PO 4 , 100 mM NaCl, 0.1% Tween-20, pH 7.5) for 1 h, the membranes were probed overnight at 4 °C with one of the following primary antibodies: rabbit polyclonal anti-AQP2 (#AQP-002, Alomone Labs, Jerusalem, Israel), rabbit polyclonal anti-pSer256-AQP2 (#ab111346, Abcam, Cambridge, MA, USA), rabbit polyclonal anti-CREB-1 (#06-863, Sigma-Aldrich), mouse monoclonal anti-p-CREB-1 (#sc-81486, Santa Cruz Biotechnology, Dallas, TX, USA), and mouse monoclonal anti-β-actin (#A5441, Sigma-Aldrich).

Techniques: Expressing, Western Blot, Control, Immunohistochemistry, Labeling, Standard Deviation, MANN-WHITNEY

Effects of duloxetine treatment on mRNA levels of aquaporin-2 (AQP2) and vasopressin-2 receptor (V2R) in lithium-induced nephrogenic diabetes insipidus. Quantitative PCR analyses of AQP2 ( A ) and V2R ( B ) were performed using whole kidneys. Compared with controls, the mRNA levels of AQP2 and V2R were decreased by lithium treatment (Li) and increased by lithium/duloxetine co-treatment (Li + DX). Each bar represents the mean ± standard deviation. *, p < 0.05 vs. control; # , p < 0.05 vs. Li by the post hoc Mann–Whitney U test.

Journal: Life

Article Title: Duloxetine-Induced Antidiuresis in Rats with Lithium-Induced Nephrogenic Diabetes Insipidus

doi: 10.3390/life14081012

Figure Lengend Snippet: Effects of duloxetine treatment on mRNA levels of aquaporin-2 (AQP2) and vasopressin-2 receptor (V2R) in lithium-induced nephrogenic diabetes insipidus. Quantitative PCR analyses of AQP2 ( A ) and V2R ( B ) were performed using whole kidneys. Compared with controls, the mRNA levels of AQP2 and V2R were decreased by lithium treatment (Li) and increased by lithium/duloxetine co-treatment (Li + DX). Each bar represents the mean ± standard deviation. *, p < 0.05 vs. control; # , p < 0.05 vs. Li by the post hoc Mann–Whitney U test.

Article Snippet: After being blocked with 5% skim milk in PBS-T (80 mM Na 2 HPO 4 , 20 mM NaH 2 PO 4 , 100 mM NaCl, 0.1% Tween-20, pH 7.5) for 1 h, the membranes were probed overnight at 4 °C with one of the following primary antibodies: rabbit polyclonal anti-AQP2 (#AQP-002, Alomone Labs, Jerusalem, Israel), rabbit polyclonal anti-pSer256-AQP2 (#ab111346, Abcam, Cambridge, MA, USA), rabbit polyclonal anti-CREB-1 (#06-863, Sigma-Aldrich), mouse monoclonal anti-p-CREB-1 (#sc-81486, Santa Cruz Biotechnology, Dallas, TX, USA), and mouse monoclonal anti-β-actin (#A5441, Sigma-Aldrich).

Techniques: Real-time Polymerase Chain Reaction, Standard Deviation, Control, MANN-WHITNEY

Reversal of duloxetine effects by tolvaptan co-treatment in lithium-induced nephrogenic diabetes insipidus. Immunoblots of AQP2, pS256-AQP2, CREB-1, and pCREB-1 are shown from the renal cortex ( A ) and medulla ( B ); each lane was loaded with a protein sample from a different rat kidney. Densitometric analysis revealed that the protein levels of AQP2, pS256-AQP2, and pCREB-1 were significantly reduced by lithium treatment (Li) compared to controls, but were restored by lithium/duloxetine co-treatment (Li + DX). However, all changes in both the cortex ( C ) and medulla ( D ) were reversed by the addition of tolvaptan (Li + DX + TV). Each group contained six rats, and the densitometry results are presented as the mean ± standard deviation. *, p < 0.05 vs. control; # , p < 0.05 vs. Li; † , p < 0.05 vs. Li + DX by the post hoc Mann–Whitney U test.

Journal: Life

Article Title: Duloxetine-Induced Antidiuresis in Rats with Lithium-Induced Nephrogenic Diabetes Insipidus

doi: 10.3390/life14081012

Figure Lengend Snippet: Reversal of duloxetine effects by tolvaptan co-treatment in lithium-induced nephrogenic diabetes insipidus. Immunoblots of AQP2, pS256-AQP2, CREB-1, and pCREB-1 are shown from the renal cortex ( A ) and medulla ( B ); each lane was loaded with a protein sample from a different rat kidney. Densitometric analysis revealed that the protein levels of AQP2, pS256-AQP2, and pCREB-1 were significantly reduced by lithium treatment (Li) compared to controls, but were restored by lithium/duloxetine co-treatment (Li + DX). However, all changes in both the cortex ( C ) and medulla ( D ) were reversed by the addition of tolvaptan (Li + DX + TV). Each group contained six rats, and the densitometry results are presented as the mean ± standard deviation. *, p < 0.05 vs. control; # , p < 0.05 vs. Li; † , p < 0.05 vs. Li + DX by the post hoc Mann–Whitney U test.

Article Snippet: After being blocked with 5% skim milk in PBS-T (80 mM Na 2 HPO 4 , 20 mM NaH 2 PO 4 , 100 mM NaCl, 0.1% Tween-20, pH 7.5) for 1 h, the membranes were probed overnight at 4 °C with one of the following primary antibodies: rabbit polyclonal anti-AQP2 (#AQP-002, Alomone Labs, Jerusalem, Israel), rabbit polyclonal anti-pSer256-AQP2 (#ab111346, Abcam, Cambridge, MA, USA), rabbit polyclonal anti-CREB-1 (#06-863, Sigma-Aldrich), mouse monoclonal anti-p-CREB-1 (#sc-81486, Santa Cruz Biotechnology, Dallas, TX, USA), and mouse monoclonal anti-β-actin (#A5441, Sigma-Aldrich).

Techniques: Western Blot, Standard Deviation, Control, MANN-WHITNEY

Renal aquaporin (AQP)2 levels in taurine-treated and taurine-deficient rats. The 25 kD and the 30-37 kD bands represent non-glycosylated and glycosylated forms of AQP2, respectively. Protein expression is expressed as percent of the control group. Data are means ± SEM (n=3 animals/groups). * p<0.05 compared to the other two groups.

Journal: Journal of Biomedical Science

Article Title: Differential effects of taurine treatment and taurine deficiency on the outcome of renal ischemia reperfusion injury

doi: 10.1186/1423-0127-17-S1-S32

Figure Lengend Snippet: Renal aquaporin (AQP)2 levels in taurine-treated and taurine-deficient rats. The 25 kD and the 30-37 kD bands represent non-glycosylated and glycosylated forms of AQP2, respectively. Protein expression is expressed as percent of the control group. Data are means ± SEM (n=3 animals/groups). * p<0.05 compared to the other two groups.

Article Snippet: Immunohistochemical staining was carried out utilizing aquaporin (AQP) 1 and AQP2 antibodies (rabbit-polyclonal antibodies to AQP1 and AQP2 (Sigma, St. Louis MO) [ , ].

Techniques: Expressing

Immunohistochemical localization of renal AQP1 and AQP2. Panel A shows immunostaining for AQP1 antibody (arrows) while panel B shows immunostaining for AQP 2 in renal tissue of TT/IR group.

Journal: Journal of Biomedical Science

Article Title: Differential effects of taurine treatment and taurine deficiency on the outcome of renal ischemia reperfusion injury

doi: 10.1186/1423-0127-17-S1-S32

Figure Lengend Snippet: Immunohistochemical localization of renal AQP1 and AQP2. Panel A shows immunostaining for AQP1 antibody (arrows) while panel B shows immunostaining for AQP 2 in renal tissue of TT/IR group.

Article Snippet: Immunohistochemical staining was carried out utilizing aquaporin (AQP) 1 and AQP2 antibodies (rabbit-polyclonal antibodies to AQP1 and AQP2 (Sigma, St. Louis MO) [ , ].

Techniques: Immunohistochemical staining, Immunostaining