aqp2 goat polyclonal antibody  (Santa Cruz Biotechnology)

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

Journal: bioRxiv

Article Title: Kidney cystogenesis in embryonic- and adult-onset ADPKD is suppressed from lack of adenylyl cyclase targeting to cilia

doi: 10.1101/2025.05.27.656198

Figure Lengend Snippet: (A) H&E images of kidneys in control, HoxB7-Cre; Pkd1 f/f , HoxB7-Cre; Pkd1 f/f ; Ankmy2 f/f , HoxB7-Cre; Ankmy2 f/f mice at P3 in C57BL/6J background. Images of multiple kidneys shown in Figure S2B. (B) 2-Kidney to body weight ratios at P3 in HoxB7-Cre; Pkd1 f/f mice were significantly higher than HoxB7-Cre; Pkd1 f/f ; Ankmy2 f/ . P3 animals had body weight between 2.5-3 g. (C) Significantly increased cystic index in HoxB7-Cre; Pkd1 f/f mice compared to HoxB7-Cre; Pkd1 f/f ; Ankmy2 f/f . (D-E) AQP2/LTL co-staining in kidney sections at P3 shows multiple Aqp2 positive cysts in medullar and corticomedullar junctions in HoxB7-Cre; Pkd1 f/f animals compared to HoxB7-Cre; Pkd1 f/f ; Ankmy2 f/f . Scale: 200 µm. Magnified images are shown in E. LTL positive, green outlines; Aqp2 positive, magenta outlines. Scale: 200 µm (D), 100 µm (E). (F) Transcript levels of Lnc2 in whole kidneys from HoxB7-Cre; Pkd1 f/f mice are significantly higher than HoxB7-Cre; Pkd1 f/f ; Ankmy2 f/f . See also Supplemental Figure 1.

Article Snippet: Sections were incubated with primary antibodies against the following antigens; overnight at room temperature or 4C: Acetylated tubulin (T6793; Sigma mouse IgG2b, 1:500), AQP2 (SC515770, Santa Cruz Biotechnology, mouse IgG1, 1:500), Ki67 (ab16667, Abcam, 1:500), pCREB (9198S; Cell Signaling, 1:500).

Techniques: Control, Staining

Journal: bioRxiv

Article Title: Kidney cystogenesis in embryonic- and adult-onset ADPKD is suppressed from lack of adenylyl cyclase targeting to cilia

doi: 10.1101/2025.05.27.656198

Figure Lengend Snippet: ( A ) Kaplan-Meier survival curves of control (N=77), HoxB7-Cre; Pkd1 f/f (N=41), HoxB7-Cre; Pkd1 f/f ; Ankmy2 f/f (N=37) and HoxB7-Cre; Ankmy2 f/f (N=37) mice. **, p=0024 by log-rank (Mantel-Cox) test. (B) H&E and whole mount images of P15 kidneys in control, HoxB7-Cre; Pkd1 f/f , HoxB7-Cre; Pkd1 f/f ; Ankmy2 f/f , HoxB7-Cre; Ankmy2 f/f mice at P15 in C57BL/6J background. Images of multiple kidneys shown in Figure S3. (C) 2-Kidney to body weight ratios at P15 in HoxB7-Cre; Pkd1 f/f mice were not significantly different from HoxB7-Cre; Pkd1 f/f ; Ankmy2 f/f . Both males (black) and females (red) are shown. (D) At P15 cystic index in HoxB7-Cre; Pkd1 f/f kidneys compared to HoxB7-Cre; Pkd1 f/f ; Ankmy2 f/f were not significantly different. (E) AQP2/pCREB co-staining in kidney sections at P15 shows comparable nuclear pCREB levels in HoxB7-Cre; Pkd1 f/f animals and HoxB7-Cre; Pkd1 f/f ; Ankmy2 f/f cysts. Scale: 50 µm. See also Supplemental Figure 2.

Article Snippet: Sections were incubated with primary antibodies against the following antigens; overnight at room temperature or 4C: Acetylated tubulin (T6793; Sigma mouse IgG2b, 1:500), AQP2 (SC515770, Santa Cruz Biotechnology, mouse IgG1, 1:500), Ki67 (ab16667, Abcam, 1:500), pCREB (9198S; Cell Signaling, 1:500).

Techniques: Control, Staining

Journal: bioRxiv

Article Title: Kidney cystogenesis in embryonic- and adult-onset ADPKD is suppressed from lack of adenylyl cyclase targeting to cilia

doi: 10.1101/2025.05.27.656198

Figure Lengend Snippet: (A) Scheme for inducible conditional knockout in kidney nephron epithelia in an adult-onset model of PKD. (B) H&E and whole mount images of 5-month-old kidneys in control, Pax8 rtTA ; TetO-Cre ; Pkd1 f/f , Pax8 rtTA ; TetO-Cre ; Pkd1 f/f ; Ankmy2 f/f , Pax8 rtTA ; TetO-Cre ; Ankmy2 f/f male mice in C57BL/6J background. Images of multiple kidneys in male mice are shown in Figure S3. (C) 2-Kidney to body weight ratios of 5-month-old Pax8 rtTA ; TetO-Cre ; Pkd1 f/f male mice were significantly higher than Pax8 rtTA ; TetO-Cre ; Pkd1 f/f ; Ankmy2 f/ . (D) Significantly increased cystic index in Pax8 rtTA ; TetO-Cre ; Pkd1 f/f male mice compared to Pax8 rtTA ; TetO-Cre ; Pkd1 f/f ; Ankmy2 f/f . (E) AQP2/LTL co-staining in kidney sections at 5 months shows multiple LTL positive cysts in cortical regions and corticomedullar junctions and AQP2 positive cysts in medullar, corticomedullar junctions and cortical regions in Pax8 rtTA ; TetO-Cre ; Pkd1 f/f animals compared to Pax8 rtTA ; TetO-Cre ; Pkd1 f/f ; Ankmy2 f/f . Scale: 300 µm. Magnified images of cortical and cortico-medullar junctions are shown below; LTL positive, green outlines; AQP2 positive, magenta outlines. Scale: 100 µm. (F) Cyst sizes in Pax8 rtTA ; TetO-Cre ; Pkd1 f/f were increased in both LTL+ and AQP2+ cysts compared to Pax8 rtTA ; TetO-Cre ; Pkd1 f/f ; Ankmy2 f/f but more so in LTL+ collecting duct cysts, which showed more differences between the two genotypes. Superplots of N=3 kidneys/genotype are shown with different shapes and averages are plotted with larger shapes. Cystic indices in Pax8 rtTA ; TetO-Cre ; Pkd1 f/f were 63, 47, and 40, whereas that in Pax8 rtTA ; TetO-Cre ; Pkd1 f/f ; Ankmy2 f/f were 22, 15, and 18. (G) The BUN values of Pax8 rtTA ; TetO-Cre ; Pkd1 f/f was significantly higher than Pax8 rtTA ; TetO-Cre ; Pkd1 f/f ; Ankmy2 f/f mice, and the levels correlated with cystic index across genotypes. See also Supplemental Figure 3 and 4.

Article Snippet: Sections were incubated with primary antibodies against the following antigens; overnight at room temperature or 4C: Acetylated tubulin (T6793; Sigma mouse IgG2b, 1:500), AQP2 (SC515770, Santa Cruz Biotechnology, mouse IgG1, 1:500), Ki67 (ab16667, Abcam, 1:500), pCREB (9198S; Cell Signaling, 1:500).

Techniques: Knock-Out, Control, Staining

Journal: bioRxiv

Article Title: Kidney cystogenesis in embryonic- and adult-onset ADPKD is suppressed from lack of adenylyl cyclase targeting to cilia

doi: 10.1101/2025.05.27.656198

Figure Lengend Snippet: (A-C) Increased proliferation in cystic kidneys in 5-month-old Pax8 rtTA ; TetO-Cre ; Pkd1 f/f male mice compared to Pax8 rtTA ; TetO-Cre ; Pkd1 f/f ; Ankmy2 f/f . Kidney sections were immunostained for Ki67, AQP2 and LTL and counterstained with DAPI. Ki67 + cells were quantified. Regression analysis with respect to cystic index is shown. (D) Immunoblotting for pERK and ERK, of (i) control, Pax8 rtTA ; (ii) TetO-Cre ; Pkd1 f/f , Pax8 rtTA ; TetO-Cre ; (iii) Pkd1 f/f ; Ankmy2 f/f and Pax8 rtTA ; (iv) TetO-Cre ; Ankmy2 f/f kidneys from male mice at 5 months. Loading controls (Tubulin) for each phosphoprotein is shown. Levels of individual phosphoprotein or total protein, each normalized to tubulin separately, is shown. Control, N=4; Pax8 rtTA ; TetO-Cre ; Pkd1 f/f N=6; Pkd1 f/f ; Ankmy2 f/f and Pax8 rtTA N=6; and TetO-Cre ; Ankmy2 f/f N=4. (E) Transcript levels of Kim1 in whole kidneys from Pax8 rtTA ; TetO-Cre ; Pkd1 f/f are significantly higher than Pax8 rtTA ; TetO-Cre ; Pkd1 f/f ; Ankmy2 f/f , and the levels correlated with cystic index across genotypes.

Article Snippet: Sections were incubated with primary antibodies against the following antigens; overnight at room temperature or 4C: Acetylated tubulin (T6793; Sigma mouse IgG2b, 1:500), AQP2 (SC515770, Santa Cruz Biotechnology, mouse IgG1, 1:500), Ki67 (ab16667, Abcam, 1:500), pCREB (9198S; Cell Signaling, 1:500).

Techniques: Western Blot, Control

Journal: bioRxiv

Article Title: Kidney cystogenesis in embryonic- and adult-onset ADPKD is suppressed from lack of adenylyl cyclase targeting to cilia

doi: 10.1101/2025.05.27.656198

Figure Lengend Snippet: (A) Kidney sections of 5-month-old mice (as in ) were immunostained for acetylated tubulin, AQP2 and LTL and counterstained with DAPI. Scale 10 μm. (B) Cilia lengths were quantified and super plots of N=2-6 male 5-month-old mice shown for each genotype. Lengths from each kidney is shown with different shapes and averages are plotted with larger shapes. Data for female mice is shown in Supplemental Figure 6A. (C) Mean cilia lengths with respect to cystic indices shown for 5-month-old mice. Cilia length in Pax8 rtTA ; TetO-Cre ; Pkd1 f/f animals scale with cystic indices but remains unchanged irrespective of cystic indices in Pax8 rtTA ; TetO-Cre ; Pkd1 f/f ; Ankmy2 f/f . Both males and females are shown. (D) Total cAMP levels with respect to cystic indices shown for respective genotypes for 5-month-old mice and the levels correlated with cystic index across genotypes irrespective of sex. (E) Scheme for inducible conditional knockout in kidney nephron epithelia in an adult-onset model of PKD to detect “pre-cystic” tubules. H&E and whole mount images of 10-week-old kidneys in control, Pax8 rtTA ; TetO-Cre ; Pkd1 f/f and Pax8 rtTA ; TetO-Cre ; Pkd1 f/f ; Ankmy2 f/f male mice in C57BL/6J background. Images of multiple kidneys, 2-kidney to body weight ratios, and cystic indices in male mice are shown in Figure S6B. (F) Cilia lengths were quantified and super plots of N=3 male 10-week-old animals shown for each genotype. Lengths from each kidney is shown with different shapes and averages are plotted with larger shapes. Immunofluorescence images are shown in Supplemental Figure 6B. See also Supplemental Figure 6.

Article Snippet: Sections were incubated with primary antibodies against the following antigens; overnight at room temperature or 4C: Acetylated tubulin (T6793; Sigma mouse IgG2b, 1:500), AQP2 (SC515770, Santa Cruz Biotechnology, mouse IgG1, 1:500), Ki67 (ab16667, Abcam, 1:500), pCREB (9198S; Cell Signaling, 1:500).

Techniques: Knock-Out, Control, Immunofluorescence