aqp2  (Bioss)

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:

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

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

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

Journal: Biology Open

Article Title: Ammonia transport mediated by urea transporter A isoforms

doi: 10.1242/bio.061655

Figure Lengend Snippet: Summary of changes in surface pH following ammonia exposure of oocytes expressing wild-type or mutant mUT-A2, mUT-A3 , or mUT-B. (A) Changes in surface pH [ΔpH S(NH3) ] of oocytes expressing wild-type mUT-A2, mUT-A3 , or mUT-B following exposure to 0.5 M NH 3 /NH 4 + . (B) Changes in surface pH [ΔpH S(NH3) ] of water-injected oocytes following exposure to 0.5 M NH 3 /NH 4 + . (C) Surface pH measurements with water-injected ( n =10), water-injected with phloretin ( n =3), UT-A2 WT ( n =6), UT-A2 with phloretin ( n =5), UT-A2 T176V ( n =5), and UT-A2 T338V ( n =6) oocytes following exposure to 0.5 M NH 3 /NH 4 + . (D) Surface pH measurements with water-injected ( n =10), water-injected with phloretin ( n =3), UT-A3 WT ( n =3), UT-A3 with phloretin ( n =3), UT-A3 T246V ( n =4), and UT-A3 T408V ( n =4) oocytes following exposure to 0.5 M NH 3 /NH 4 + . (E) Surface pH measurements with water-injected ( n =10), water-injected with phloretin ( n =3), UT-B WT ( n =4), UT-B WT with phloretin ( n =4), UT-B T172V ( n =4), and UT-B T334V ( n =4) oocytes. Panels F–H on the right display background-subtracted results that yield channel-dependent activity. Oocytes expressing RhCG ( n =5; panels I and K) or AQP2 ( n =7; panels J and L) were used as positive and negative controls for assessing NH 3 transport. Gray circles indicate the values of each oocyte. Colored circles and triangles represent the averages of experiments performed on different days. The horizontal and vertical (error bars) lines on each plot represent the overall mean and the standard error of the mean of each group and condition. A standard two-tailed Student's t -test with Bonferroni correction was used to compare the difference between two means, as indicated above each graph. The significance level was set at P <0.0125.

Article Snippet: Experiments using oocytes injected with hAQP2 or hRhCG employed the same processing and detection protocols used for the UTs, except that a polyclonal anti-AQP2 antibody (Alpha Diagnostics, San Antonio, TX, USA) or a polyclonal antibody raised against the C-terminal region of RhCG ( ) and a goat anti-rabbit secondary antibody conjugated to HRP (AP132P; Millipore, Billerica, MA, USA) were used as reported previously ( , ).

Techniques: Expressing, Mutagenesis, Injection, Activity Assay, Two Tailed Test

Journal: Journal of Translational Medicine

Article Title: ANKHD1 promotes pathogenic proliferation in Autosomal Dominant Polycystic Kidney Disease via the Cyclin D1/CDK4 pathway

doi: 10.1186/s12967-025-06359-9

Figure Lengend Snippet: Ankhd1 , a protein that controls proliferation, is expressed in epithelial cells of normal and ADPKD kidneys. Expression pattern of Ankhd1 protein in ( A ) WT and ( B ) Pkd1 nl/nl mouse kidneys. ANKHD1 shown in green. TOPRO (blue) was used for nuclear counterstaining. C ) ANKHD1 is highly expressed in cyst lining cells of Pkd1 nl/nl Ankhd1 +/+ mouse kidneys while is reduced in Ankhd1 +/− mice. D ) Quantification of ANKHD1 was performed and data are presented as ± S.E.M. Each dot represents an individual mouse kidney. Student’s t -test was used to calculate the indicated statistical significance. ANKHD1 is expressed in proximal tubules of Pkd1 nl/nl mice as shown by Aquaporin-1 (AQP1) staining ( E ), and also in collecting ducts as shown by Aquaporin-2 (AQP2) staining ( F ). Scale bars are 500 μm

Article Snippet: The antibodies that were used are: ANKHD1 Rabbit mAb (HPA008718, Sigma), p21 Waf1/Cip1 (12D1) Rabbit mAb (#2947, Cell Signaling), CDKN2D Rabbit polyclonal antibody (p19) (BS6940, Bioworld Technology), Cyclin D1 (E3P5S) XP ® Rabbit mAb (#55506, Cell Signaling), CDK4 (D9G3E) Rabbit mAb (#12790, Cell Signaling), phospho-RB, AQP1 Rabbit polyclonal antibody (Santa Cruz Biotechnology, sc-20810), AQP2 Goat polyclonal antibody (Santa Cruz Biotechnology, sc-9882), Ki67 Rabbit polyclonal antibody (ab15580, Abcam), β-actin Mouse mAb (ab8224, Abcam), goat anti-mouse IgG/HRP (P0447, Dako), goat anti-rabbit IgG/HRP (P0448, Dako).

Techniques: Expressing, Staining