Journal: Frontiers in Physiology

Article Title: Empagliflozin Contributes to Polyuria via Regulation of Sodium Transporters and Water Channels in Diabetic Rat Kidneys

doi: 10.3389/fphys.2019.00271

Figure Lengend Snippet: Empagliflozin causes no significant changes to AQP3 and AQP1 but tends to increase AQP7 level in diabetic rat kidneys. (A) Representative renal sections immunostained with anti-AQP3. (B) Quantitative analysis of results for AQP3 shows no significant change among groups. Magnification, ×200. (C) Representative immunoblot reacting with anti-NCC, anti-α-ENaC and anti- γ-ENaC. (D) Densitometric analysis shows that the expression of AQP3 protein significantly increased in all OLETF rats. Lixisenatide treatment further increased the expression of AQP3. ∗ P < 0.001 vs. other groups. † P < 0.001 vs. OLETF_C; P = 0.002 vs. OLETF_E; P = 0.003 vs. OLETF_V. (E) qRT-PCR shows that mRNA level of AQP3 significantly increased in all OLETF rats. ∗ P = 0.001 vs. OLETF_C; P < 0.001 vs. OLETF_E and OLETF_L; P = 0.001 vs. OLETF_V. (F) Densitometric analysis shows that AQP7 in kidneys was significantly lower in untreated OLETF group than in LETO group and all treated OLETF groups. ∗ P < 0.001 vs. LETO and OLETF_E; P = 0.001 vs. OLETF_L and OLETF_V. (G) Densitometric analysis shows the decreased AQP1 expressions in all OLETF rats. ∗ P < 0.001 vs. other groups. † P = 0.046 vs. OLETF_C; P < 0.001 vs. OLETF_E; P = 0.047 vs. OLETF_L (H) mRNA level of AQP1, decreased in untreated OLETF rats, was increased with antidiabetic treatment. ∗ P < 0.001 vs. LETO; P = 0.003 vs. OLETF_E; P = 0.001 vs. OLETF_L; P = 0.002 vs. OLETF_V. (I) Representative renal sections immunostained with anti-AQP1. (J) Quantitative analysis of results for AQP1 shows no significant changes among groups. Magnification, ×200. n = 8 per each group.

Article Snippet: Membranes were incubated with the following primary antibodies: AQP1 (Alomone Labs, Jerusalem, Israel), AQP2 (Alomone Labs), p261-AQP2 (Novus Biologicals, Littleton, CO, United States), AQP3 (Alomone Labs), AQP7 (Alomone Labs), cyclin-dependent kinase 1 (cdk1; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, United States), cdk5 (Santa Cruz Biotechnology, Inc.), Na+/H+ exchanger isoform 3 (NHE3; StressMarq Biosciences Inc., Victoria, BC, Canada), extracellular signal–regulated kinase (ERK; Cell Signaling Technology, Danvers, MA, United States), p-Erk (Cell Signaling Technology), p -glycogen synthase kinase 3α (pGSK3α; Santa Cruz Biotechnology, Inc.), Na+-Cl- cotransporter (NCC; StressMarq Biosciences), NKCC2 (StressMarq Biosciences), phosphatase 1β (PP1β; Santa Cruz Biotechnology, Inc.), PP2B (Santa Cruz Biotechnology, Inc.), vasopressin V2 receptor (V2R; Alomone Labs), α-epithelial Na+ channel (ENaC; StressMarq Biosciences); γ-ENaC (StressMarq Biosciences), p38 mitogen-activated protein kinase (p38; Cell Signaling Technology), p-p38 (Cell Signaling Technology) and β-actin (Sigma-Aldrich, St. Louis, MO, United States).

Techniques: Western Blot, Expressing, Quantitative RT-PCR

Journal: Cells

Article Title: Aquaglyceroporins in Human Breast Cancer

doi: 10.3390/cells12172185

Figure Lengend Snippet: Graphical illustration of workflow. Formalin-fixed, paraffin-embedded samples from invasive female breast cancer tumors were obtained in fully anonymized form from surgical specimens from the Department of Pathology, Aarhus University Hospital, Denmark. Following retrieval, the anonymized tissue samples were sectioned. For diagnostic assessment, hematoxylin and eosin (H&E) stainings, as well as immunohistochemical stainings for E-cadherin, were performed. Samples representing invasive ductal carcinoma (IDC) and invasive lobular carcinoma (ILC) were further analyzed. Serial sections were processed for immunohistochemical stainings in the following order: #1 AQP7, #2 AQP9, #3 AQP3 and #4 SMMS-1. The SMMS-1 staining identified myoepithelial cells, allowing a distinction between in situ and invasion lesions. Images of all samples were captured by brightfield light microscopy, and select slides for publication figures were scanned on a NanoZoomer 2.0HT (Hamamatsu) with the 40× objective. Part of this figure was generated in BioRender.com; accessed on August 11th 2023.

Article Snippet: The previously validated primary antibodies were: section #1 rabbit anti-AQP7 diluted 1:200 [ ], section #2 rabbit-anti-AQP9 diluted 1:50 [ , ], section #3 rabbit anti-AQP3 diluted 1:400 (Alomone Labs, Cat# AQP-003, Jerusalem, Israel) and section #4 mouse-anti-Smooth Muscle Myosin, heavy chain (SMMS-1) diluted 1:2000 (Cell Marque, Cat# 298M-15, Rocklin, CA, USA).

Techniques: Formalin-fixed Paraffin-Embedded, Diagnostic Assay, Immunohistochemistry, Staining, In Situ, Light Microscopy, Generated

Journal: Cells

Article Title: Aquaglyceroporins in Human Breast Cancer

doi: 10.3390/cells12172185

Figure Lengend Snippet: AQP7, AQP9 and AQP3 localize to epithelial cells of lobules and extralobular ducts in benign regions adjacent to tumor tissue. Formalin-fixed, paraffin-embedded samples from invasive female breast cancer tumors were obtained in fully anonymized form from surgical specimens and processed for immunohistochemistry with antibodies against AQP7, AQP9, AQP3 and SMMS-1. The figure depicts representative images from immunohistochemical stainings of serial sections from the normal/benign part of samples from invasive lobular carcinoma. ( A ) Lobule and ( B ) extralobular duct. Immune cells in the surrounding connective tissue also stain positive (red asterisks in B). Arrowheads in B point to myoepithelial cells, and the arrows in B point to the apical region of extralobular ducts. Slides were scanned in a NanoZoomer 2.0HT (Hamamatsu) using the 40× objective. White balance was adjusted in Adobe Photoshop CS4. Scale bars are 100 µm.

Article Snippet: The previously validated primary antibodies were: section #1 rabbit anti-AQP7 diluted 1:200 [ ], section #2 rabbit-anti-AQP9 diluted 1:50 [ , ], section #3 rabbit anti-AQP3 diluted 1:400 (Alomone Labs, Cat# AQP-003, Jerusalem, Israel) and section #4 mouse-anti-Smooth Muscle Myosin, heavy chain (SMMS-1) diluted 1:2000 (Cell Marque, Cat# 298M-15, Rocklin, CA, USA).

Techniques: Formalin-fixed Paraffin-Embedded, Immunohistochemistry, Staining

Journal: Cells

Article Title: Aquaglyceroporins in Human Breast Cancer

doi: 10.3390/cells12172185

Figure Lengend Snippet: AQP7, AQP9 and AQP3 localize to neoplastic cells of lobular carcinoma in situ and invasive lobular carcinoma. Formalin-fixed, paraffin-embedded samples from invasive female breast cancer tumors were obtained in fully anonymized form from surgical specimens and processed for immunohistochemistry with antibodies against AQP7, AQP9, AQP3 and SMMS-1. The figure depicts representative images from immunohistochemical stainings of serial sections from patients with invasive lobular carcinoma (ILC) and enlarged images (inserts) of the marked areas. ( A ) Images are from a region of lobular carcinoma in situ (LCIS) from an ILC patient, and ( B ) is ILC. Slides were scanned in a NanoZoomer 2.0HT (Hamamatsu) using the 40× objective. White balance was adjusted in Adobe Photoshop CS4. Scale bars are 100 µm ( A , B ) and 20 µm (( A , B ), inserts).

Article Snippet: The previously validated primary antibodies were: section #1 rabbit anti-AQP7 diluted 1:200 [ ], section #2 rabbit-anti-AQP9 diluted 1:50 [ , ], section #3 rabbit anti-AQP3 diluted 1:400 (Alomone Labs, Cat# AQP-003, Jerusalem, Israel) and section #4 mouse-anti-Smooth Muscle Myosin, heavy chain (SMMS-1) diluted 1:2000 (Cell Marque, Cat# 298M-15, Rocklin, CA, USA).

Techniques: In Situ, Formalin-fixed Paraffin-Embedded, Immunohistochemistry

Journal: Cells

Article Title: Aquaglyceroporins in Human Breast Cancer

doi: 10.3390/cells12172185

Figure Lengend Snippet: AQP7, AQP9 and AQP3 localize to neoplastic cells of ductal carcinoma in situ and invasive ductal carcinoma. Formalin-fixed, paraffin-embedded samples from invasive female breast cancer tumors were obtained in fully anonymized form from surgical specimens and processed for immunohistochemistry with antibodies against AQP7, AQP9, AQP3 and SMMS-1. The figure depicts representative images from immunohistochemical stainings of serial sections from patients with invasive ductal carcinoma (IDC) and enlarged images (inserts) of the marked areas. ( A ) Images are from a region of ductal carcinoma in situ (DCIS) from an IDC patient, and ( B ) is IDC. Slides were scanned in a NanoZoomer 2.0HT (Hamamatsu) using the 40x objective. White balance was adjusted in Adobe Photoshop CS4. Scale bars are 175 µm ( A , B ) and 50 µm (( A , B ), inserts).

Article Snippet: The previously validated primary antibodies were: section #1 rabbit anti-AQP7 diluted 1:200 [ ], section #2 rabbit-anti-AQP9 diluted 1:50 [ , ], section #3 rabbit anti-AQP3 diluted 1:400 (Alomone Labs, Cat# AQP-003, Jerusalem, Israel) and section #4 mouse-anti-Smooth Muscle Myosin, heavy chain (SMMS-1) diluted 1:2000 (Cell Marque, Cat# 298M-15, Rocklin, CA, USA).

Techniques: In Situ, Formalin-fixed Paraffin-Embedded, Immunohistochemistry