Journal: International Dental Journal

Article Title: ATP1A1-Driven Intercellular Contact Between Dental Pulp Stem Cell and Endothelial Cell Enhances Vasculogenic Activity

doi: 10.1016/j.identj.2025.100870

Figure Lengend Snippet: RNA-sequencing data and the expression of ATP1A1 and α-SMA after direct coculture. (A-B) Differently expressed Genes (DEGs) analysis of cells after direct coculture. (C-D) Gene ontology (GO) analysis revealed the enrichment of biological processes among the upregulated genes of DPSCs. (E-F) RT-qPCR analysis of the relative mRNA expression of ATP1A1, ATP1B1 and α-SMA in cells after coculture for 24 and 48 hours. (G) Representative immunofluorescence images of ATP1A1(green) and DAPI (blue) in HUVECs after coculture for 2 days. Scale bar = 20 μm. (H-I) Western blot analysis of the expression of ATP1A1 and α-SMA in DPSCs after coculture for 48 hours. Data are mean ± SD for n = 3 replicates, * P < .05, *** P < .001.

Article Snippet: hDPSCs were transfected at approximately 60% confluency in opti-mem medium (antibiotic-free, Gibco) with 50nM siRNA targeting ATP1A1 (MCE), using Lipofectamine 2000 reagent (Invitrogen, Thermo Fisher Scientific).

Techniques: RNA Sequencing, Expressing, Quantitative RT-PCR, Immunofluorescence, Western Blot

Journal: International Dental Journal

Article Title: ATP1A1-Driven Intercellular Contact Between Dental Pulp Stem Cell and Endothelial Cell Enhances Vasculogenic Activity

doi: 10.1016/j.identj.2025.100870

Figure Lengend Snippet: DPSCs exhibited differentiation properties toward SMCs after contact with HUVECs. (A) RT-qPCR analysis of the relative mRNA expression of ATP1A1, ATP1B1 and α-SMA in DPSCs after coculture for 24 and 48 hours. (B) Western blot analysis of the expression of ATP1A1, and α-SMA in DPSCs after coculture for 24 and 48 hours. (C-D) Representative immunofluorescence images of α-SMA (green) and DAPI (blue) in HUVECs after coculture for 2 days. Scale bar = 50 μm. (E-F) Microscopy images showed the location of ATP1A1 and α-SMA in direct coculture. Scale bar = 50 μm. Data are mean ± SD for n = 3 replicates, * P < .05, ** P < .01, *** P < .001.

Article Snippet: hDPSCs were transfected at approximately 60% confluency in opti-mem medium (antibiotic-free, Gibco) with 50nM siRNA targeting ATP1A1 (MCE), using Lipofectamine 2000 reagent (Invitrogen, Thermo Fisher Scientific).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Immunofluorescence, Microscopy

Journal: International Dental Journal

Article Title: ATP1A1-Driven Intercellular Contact Between Dental Pulp Stem Cell and Endothelial Cell Enhances Vasculogenic Activity

doi: 10.1016/j.identj.2025.100870

Figure Lengend Snippet: Inhibition of ATP1A1 suppresses the expression of SMC markers in DPSCs. (A-D) RT-qPCR analysis of the relative expression of ATP1A1, α-SMA and activin A in cells after siRNA transfection. Representative fluorescence images of cells after siRNA transfection. Images were captured at 48 hours post-transfection to visualise transfection efficiency and cell morphology. (E-F) Western blot analysis of the relative expression of proteins in cells after siRNA transfection. (G) The impact of ouabain treatment on DPSCs. (H-J) Ouabain inhibited HUVEC migration and tube formation. Data are mean ± SD for n = 3 replicates, * P < .05, *** P < .001, Scale Bar = 100 μm.

Article Snippet: hDPSCs were transfected at approximately 60% confluency in opti-mem medium (antibiotic-free, Gibco) with 50nM siRNA targeting ATP1A1 (MCE), using Lipofectamine 2000 reagent (Invitrogen, Thermo Fisher Scientific).

Techniques: Inhibition, Expressing, Quantitative RT-PCR, Transfection, Fluorescence, Western Blot, Migration

Journal: International Dental Journal

Article Title: ATP1A1-Driven Intercellular Contact Between Dental Pulp Stem Cell and Endothelial Cell Enhances Vasculogenic Activity

doi: 10.1016/j.identj.2025.100870

Figure Lengend Snippet: Involvement of Src/AKT pathway in HUVECs/DPSCs direct coculture. (A-B) Effects of ATP1A1 on the phosphorylation status of AKT and Src after coculture. (C-D) Effects of ATP1A1 on the phosphorylation status of AKT and Src in DPSCs. (E-F) Semi-quantitative analysis of the phosphorylation and total protein after coculture. Data are mean ± SD for n = 3 replicates, * P < .05, ** P < .01.

Article Snippet: hDPSCs were transfected at approximately 60% confluency in opti-mem medium (antibiotic-free, Gibco) with 50nM siRNA targeting ATP1A1 (MCE), using Lipofectamine 2000 reagent (Invitrogen, Thermo Fisher Scientific).

Techniques: Phospho-proteomics

Journal: International Dental Journal

Article Title: ATP1A1-Driven Intercellular Contact Between Dental Pulp Stem Cell and Endothelial Cell Enhances Vasculogenic Activity

doi: 10.1016/j.identj.2025.100870

Figure Lengend Snippet: Proposed model of cell interactions through ATP1A1 promotes SMC activation for vascular stability. Cell interactions between DPSCs and HUVECs induce ATP1A1 over-expression, leading to SMC differentiation, Src/AKT activation and activin A, FGF-2 secretion. The schematic diagram of mechanism was generated by Figdraw.

Article Snippet: hDPSCs were transfected at approximately 60% confluency in opti-mem medium (antibiotic-free, Gibco) with 50nM siRNA targeting ATP1A1 (MCE), using Lipofectamine 2000 reagent (Invitrogen, Thermo Fisher Scientific).

Techniques: Activation Assay, Over Expression, Generated

Journal: Frontiers in Medicine

Article Title: Construction of tissue engineered cornea with skin-derived corneal endothelial-like cell and mechanism research for the cell differentiation

doi: 10.3389/fmed.2024.1448248

Figure Lengend Snippet: SKPs differentiated into CEC-like cells. (A) SKPs were cultured as floating spheres. During cell differentiation, the morphology of the cells changed gradually. On day 10, the cells became the most endothelial-like and formed a mosaic monolayer. (B) Immunofluorescence showed that the CEC-like cells expressed CEC markers Na + /K + ATPase, ZO-1, and Pitx2 after 10 days of differentiation. (C) RT-PCR showed that the CEC-like cells expressed CEC markers Pax6, Cdh2, Car2, Slc4a4, Col4a2 and Col8a2. (Data are mean ± SEM, * p < 0.05, n = 3, scale bar = 100 μm).

Article Snippet: The primary antibodies used included Na + /K + ATPase (1:100, Novus, NB300-146), ZO-1 (1:100, CST, 13663S), PITX2 (1:100, Abcam, ab98297), and human nuclei (1,100, Millipore, MAB1281).

Techniques: Cell Culture, Cell Differentiation, Immunofluorescence, Reverse Transcription Polymerase Chain Reaction

Journal: International journal of molecular sciences

Article Title: A Comparative Study on the Lysosomal Cation Channel TMEM175 Using Automated Whole-Cell Patch-Clamp, Lysosomal Patch-Clamp, and Solid Supported Membrane-Based Electrophysiology: Functional Characterization and High-Throughput Screening Assay Development.

doi: 10.3390/ijms241612788

Figure Lengend Snippet: Figure 1. Principles of SSM-based electrophysiology recordings on TMEM175 localized in lysosomes: (A) Schematic of the steps performed to purify lysosomes from HEK293 cell culture. The process is based on a protocol by Schulz et al. [38]; (B) Marker protein concentrations for lysosomes (LAMP-1) and plasma membrane (Na-K-ATPase) for lysosomal samples purified according to Schulz et al. [38] and Jinn et al. [5] were determined via ELISA. The graph shows the amount of marker proteins nor- malized to the total protein amount of the respective sample determined via Bradford assay. Average values and standard deviations from N = 2 samples are shown; (C) Schematic of lysosomes adsorbed to the SSM on a gold-coated sensor chip. The capacitive read-out is highlighted; (D) SURFE2R N1 platform for SSME recordings in a single-well format; (E) Three-millimeter sensor for the recordings with the SURFE2R N1; (F) Screenshot of the SURFE2R N1 control 1.7.0.2 software; (G) Representative current traces recorded with TMEM175 overexpressing lysosomes (blue trace) and control lysosomes (red trace) using the SURFE2R N1. The sensors harboring the TMEM175 and the control samples were each loaded with 2.2 µg total protein. SD and average current amplitudes and time constants determined from N = 6 sensors are shown in Table 1. The experiment shows a single solution ex- change from a solution containing 50 mM Na+ (NA solution, red bars) to a solution containing 50 mM K+ (A solution, green bar), which stimulates K+ flux through TMEM175 (on-signal). After 1 s, K+

Article Snippet: To determine the efficacy of fractionation and membrane enrichment procedures, the lysosomal marker LAMP1 and the plasma membrane marker Na-K-ATPase were quantified using quantitative sandwich ELISA kits (Human Sodium/Potassium-Transporting ATPase Subunit Alpha-1(ATP1A1) ELISA Kit, Cusabio, Houston, TX, USA, CSB-EL002322HU; Human LAMP1 ELISA Kit, Biorbyt, Cambridge, UK, Orb565022) according to the manufacturer’s instructions.

Techniques: Cell Culture, Marker, Clinical Proteomics, Membrane, Enzyme-linked Immunosorbent Assay, Bradford Assay, Control, Software

Journal: Cell and tissue research

Article Title: "Reversed polarization" of Na/K-ATPase-a sign of inverted transport in the human endolymphatic sac: a super-resolution structured illumination microscopy (SR-SIM) study.

doi: 10.1007/s00441-019-03106-7

Figure Lengend Snippet: Fig. 2 Immunohistochemistry of a guinea pig choroid plexus (a, b) and a human ES obtained at surgery (c). Na/K-ATPase α1 and β3 subunits are expressed in the apical cell membrane (a). Framed area is magnified in b. The apical cell membrane (small arrow) ex- presses the α1 subunit, while the β3 isoform is strongly expressed in the perinuclear zone (long ar- row). An ES was labeled with antibodies against both β1 and β3 isoforms from different species (c). In the proximal part of the sac (left), the β1 isoform was more expressed and in the basolateral cell membrane of the epithelium. In the distal part (right), the β3 subunit was more expressed and in the apical cell membranes in the epithelium. Here, only a few cells express the β1 isoform (in- set). Left frame is magnified in Fig. 3

Article Snippet: At high magnification, Na/K-ATPase protein was seen to translocate from organelles near the cell nucleus to *α1 subunit mostly co-expressed with β3 subunit Table 1 Antibodies used in the study Antibody Type Dilution Host Catalog number Producer Na/K-ATPase β1 Monoclonal 1:100 Mouse MA3-930 Thermo Fisher, Waltham, USA Na/K-ATPase β2 Polyclonal 1:50 Rabbit ANP-012 Alomone Labs, Jerusalem, Israel Na/K-ATPase β3 Polyclonal 1:200 Rabbit Abx111158 Abbexa, Cambridge, UK Na/K-ATPase α1′ Monoclonal 1:50 Mouse SC 21712 Santa Cruz Biotechnology, Dallas, USA Na/K-ATPase α1′′ Monoclonal 1:50 Mouse NB300-146 Novus, Littleton, USA Na/K-ATPase α2 Polyclonal 1:100 Rabbit OAAB03232 Aviva Systems Biology, San Diego, USA Na/K-ATPase α3 Monoclonal 1:50 Mouse NB300-540 Novus, Littleton, USA Carbonic anhydrase II Polyclonal 1:50 Rabbit HPA001550 Atlas Antibodies, Stockholm, Sweden NKCC2 Polyclonal 1:200 Rabbit OABB01331 Aviva Systems Biology, San Diego, USA Mineralocorticoid receptor′ Monoclonal 1:50 Mouse MA1-620 Thermo Fisher, Waltham, USA Mineralocorticoid receptor′′ Polyclonal 1:200 Rabbit PA5-81527 Thermo Fisher, Waltham, USA Vasopressin receptor type 2 Polyclonal 1:50 Rabbit Abx133128 Abbexa, Cambridge, UK ENaC Polyclonal 1:100 Rabbit PA1-920A Thermo Fisher, Waltham, USA Aquaporin-2 Polyclonal 1:50 Rabbit NB110-74682 Novus, Littleton, USA Aquaporin-4 Polyclonal 1:50 Rabbit AQP-004 Alomone Labs, Jerusalem, Israel Pendrin Polyclonal 1:50 Rabbit NBP1-60106 Novus, Littleton, USA NKCC1 Polyclonal 1:100 Rabbit AB59791 Abcam, Cambridge, UK NCC Polyclonal 1:100 Rabbit PA5-77816 Thermo Fisher, Waltham, USA the lateral plasma membrane (Fig. 4, insets).

Techniques: Immunohistochemistry, Membrane, Labeling

Journal: Cell and tissue research

Article Title: "Reversed polarization" of Na/K-ATPase-a sign of inverted transport in the human endolymphatic sac: a super-resolution structured illumination microscopy (SR-SIM) study.

doi: 10.1007/s00441-019-03106-7

Figure Lengend Snippet: Fig. 3 Expression of Na/K-ATPase β1 isoform (SIM, single optical sec- tion) in the epithelium of the intraosseous part of the human ES. The basolateral cell membranes strongly express the transporter protein. Arrows mark an epithelial protrusion into the sac lumen (Lu). A capillary (cap) is seen in the loose connective tissue. Inset shows a cell expressing Na/K-ATPase at higher magnification

Article Snippet: At high magnification, Na/K-ATPase protein was seen to translocate from organelles near the cell nucleus to *α1 subunit mostly co-expressed with β3 subunit Table 1 Antibodies used in the study Antibody Type Dilution Host Catalog number Producer Na/K-ATPase β1 Monoclonal 1:100 Mouse MA3-930 Thermo Fisher, Waltham, USA Na/K-ATPase β2 Polyclonal 1:50 Rabbit ANP-012 Alomone Labs, Jerusalem, Israel Na/K-ATPase β3 Polyclonal 1:200 Rabbit Abx111158 Abbexa, Cambridge, UK Na/K-ATPase α1′ Monoclonal 1:50 Mouse SC 21712 Santa Cruz Biotechnology, Dallas, USA Na/K-ATPase α1′′ Monoclonal 1:50 Mouse NB300-146 Novus, Littleton, USA Na/K-ATPase α2 Polyclonal 1:100 Rabbit OAAB03232 Aviva Systems Biology, San Diego, USA Na/K-ATPase α3 Monoclonal 1:50 Mouse NB300-540 Novus, Littleton, USA Carbonic anhydrase II Polyclonal 1:50 Rabbit HPA001550 Atlas Antibodies, Stockholm, Sweden NKCC2 Polyclonal 1:200 Rabbit OABB01331 Aviva Systems Biology, San Diego, USA Mineralocorticoid receptor′ Monoclonal 1:50 Mouse MA1-620 Thermo Fisher, Waltham, USA Mineralocorticoid receptor′′ Polyclonal 1:200 Rabbit PA5-81527 Thermo Fisher, Waltham, USA Vasopressin receptor type 2 Polyclonal 1:50 Rabbit Abx133128 Abbexa, Cambridge, UK ENaC Polyclonal 1:100 Rabbit PA1-920A Thermo Fisher, Waltham, USA Aquaporin-2 Polyclonal 1:50 Rabbit NB110-74682 Novus, Littleton, USA Aquaporin-4 Polyclonal 1:50 Rabbit AQP-004 Alomone Labs, Jerusalem, Israel Pendrin Polyclonal 1:50 Rabbit NBP1-60106 Novus, Littleton, USA NKCC1 Polyclonal 1:100 Rabbit AB59791 Abcam, Cambridge, UK NCC Polyclonal 1:100 Rabbit PA5-77816 Thermo Fisher, Waltham, USA the lateral plasma membrane (Fig. 4, insets).

Techniques: Expressing

Journal: Cell and tissue research

Article Title: "Reversed polarization" of Na/K-ATPase-a sign of inverted transport in the human endolymphatic sac: a super-resolution structured illumination microscopy (SR-SIM) study.

doi: 10.1007/s00441-019-03106-7

Figure Lengend Snippet: Fig. 4 Nanoscopic resolution of Na/K-ATPase β1 isoform and CA ex- pression (single optical section) in a cylindrical cell the intermediate por- tion of the human ES. Inset showing level of sectioning (*) of the ES. There is a strong expression of Na/K-ATPase in the lateral/basolateral cell membrane (a). A part of the apical membrane also seems to express the β isoform (arrow). Higher magnification of protein aggregates in the cyto- sol bridging the space between the cell nucleus (Nu) and the lateral plas- ma membrane (b, c)

Article Snippet: At high magnification, Na/K-ATPase protein was seen to translocate from organelles near the cell nucleus to *α1 subunit mostly co-expressed with β3 subunit Table 1 Antibodies used in the study Antibody Type Dilution Host Catalog number Producer Na/K-ATPase β1 Monoclonal 1:100 Mouse MA3-930 Thermo Fisher, Waltham, USA Na/K-ATPase β2 Polyclonal 1:50 Rabbit ANP-012 Alomone Labs, Jerusalem, Israel Na/K-ATPase β3 Polyclonal 1:200 Rabbit Abx111158 Abbexa, Cambridge, UK Na/K-ATPase α1′ Monoclonal 1:50 Mouse SC 21712 Santa Cruz Biotechnology, Dallas, USA Na/K-ATPase α1′′ Monoclonal 1:50 Mouse NB300-146 Novus, Littleton, USA Na/K-ATPase α2 Polyclonal 1:100 Rabbit OAAB03232 Aviva Systems Biology, San Diego, USA Na/K-ATPase α3 Monoclonal 1:50 Mouse NB300-540 Novus, Littleton, USA Carbonic anhydrase II Polyclonal 1:50 Rabbit HPA001550 Atlas Antibodies, Stockholm, Sweden NKCC2 Polyclonal 1:200 Rabbit OABB01331 Aviva Systems Biology, San Diego, USA Mineralocorticoid receptor′ Monoclonal 1:50 Mouse MA1-620 Thermo Fisher, Waltham, USA Mineralocorticoid receptor′′ Polyclonal 1:200 Rabbit PA5-81527 Thermo Fisher, Waltham, USA Vasopressin receptor type 2 Polyclonal 1:50 Rabbit Abx133128 Abbexa, Cambridge, UK ENaC Polyclonal 1:100 Rabbit PA1-920A Thermo Fisher, Waltham, USA Aquaporin-2 Polyclonal 1:50 Rabbit NB110-74682 Novus, Littleton, USA Aquaporin-4 Polyclonal 1:50 Rabbit AQP-004 Alomone Labs, Jerusalem, Israel Pendrin Polyclonal 1:50 Rabbit NBP1-60106 Novus, Littleton, USA NKCC1 Polyclonal 1:100 Rabbit AB59791 Abcam, Cambridge, UK NCC Polyclonal 1:100 Rabbit PA5-77816 Thermo Fisher, Waltham, USA the lateral plasma membrane (Fig. 4, insets).

Techniques: Expressing, Membrane

Journal: Cell and tissue research

Article Title: "Reversed polarization" of Na/K-ATPase-a sign of inverted transport in the human endolymphatic sac: a super-resolution structured illumination microscopy (SR-SIM) study.

doi: 10.1007/s00441-019-03106-7

Figure Lengend Snippet: Fig. 6 SR-SIM (single optical section) of human ES (intermediate portion). Inset showing level of sectioning (*) of the ES. Expression of Na/K-ATPase α1 and β3 isoforms (a). There is strong co-labeling at the apical cell membrane in some epithelial cells (arrow at inset). High mag- nification of the apical cell membrane in a cell displayed in the framed area in c (b). The β3 isoform is also expressed in the cell nucleus (arrows) and apical cytoplasm near the cell membrane. Inset in c shows nanoscopic resolution of ion transporter isoforms in the apical cell membrane

Article Snippet: At high magnification, Na/K-ATPase protein was seen to translocate from organelles near the cell nucleus to *α1 subunit mostly co-expressed with β3 subunit Table 1 Antibodies used in the study Antibody Type Dilution Host Catalog number Producer Na/K-ATPase β1 Monoclonal 1:100 Mouse MA3-930 Thermo Fisher, Waltham, USA Na/K-ATPase β2 Polyclonal 1:50 Rabbit ANP-012 Alomone Labs, Jerusalem, Israel Na/K-ATPase β3 Polyclonal 1:200 Rabbit Abx111158 Abbexa, Cambridge, UK Na/K-ATPase α1′ Monoclonal 1:50 Mouse SC 21712 Santa Cruz Biotechnology, Dallas, USA Na/K-ATPase α1′′ Monoclonal 1:50 Mouse NB300-146 Novus, Littleton, USA Na/K-ATPase α2 Polyclonal 1:100 Rabbit OAAB03232 Aviva Systems Biology, San Diego, USA Na/K-ATPase α3 Monoclonal 1:50 Mouse NB300-540 Novus, Littleton, USA Carbonic anhydrase II Polyclonal 1:50 Rabbit HPA001550 Atlas Antibodies, Stockholm, Sweden NKCC2 Polyclonal 1:200 Rabbit OABB01331 Aviva Systems Biology, San Diego, USA Mineralocorticoid receptor′ Monoclonal 1:50 Mouse MA1-620 Thermo Fisher, Waltham, USA Mineralocorticoid receptor′′ Polyclonal 1:200 Rabbit PA5-81527 Thermo Fisher, Waltham, USA Vasopressin receptor type 2 Polyclonal 1:50 Rabbit Abx133128 Abbexa, Cambridge, UK ENaC Polyclonal 1:100 Rabbit PA1-920A Thermo Fisher, Waltham, USA Aquaporin-2 Polyclonal 1:50 Rabbit NB110-74682 Novus, Littleton, USA Aquaporin-4 Polyclonal 1:50 Rabbit AQP-004 Alomone Labs, Jerusalem, Israel Pendrin Polyclonal 1:50 Rabbit NBP1-60106 Novus, Littleton, USA NKCC1 Polyclonal 1:100 Rabbit AB59791 Abcam, Cambridge, UK NCC Polyclonal 1:100 Rabbit PA5-77816 Thermo Fisher, Waltham, USA the lateral plasma membrane (Fig. 4, insets).

Techniques: Expressing, Labeling, Membrane

Journal: Cell and tissue research

Article Title: "Reversed polarization" of Na/K-ATPase-a sign of inverted transport in the human endolymphatic sac: a super-resolution structured illumination microscopy (SR-SIM) study.

doi: 10.1007/s00441-019-03106-7

Figure Lengend Snippet: Fig. 5 SR-SIM (single optical section) of an epithelial villus in the human ES (intermediate portion). Inset showing level of sectioning (*) of the ES. Expression of Na/K-ATPase α1 isoform and epithelial sodium channel (ENaC). b. Strong expression of Na/K-ATPase is localized only to the apical cell membrane. c. Nanoscopic resolution shows molecular relation- ship between Na/K-ATPase and ENAC in the apical cytoplasm. d. ENAC fluorescence at the juxta-nuclear region (arrow)

Article Snippet: At high magnification, Na/K-ATPase protein was seen to translocate from organelles near the cell nucleus to *α1 subunit mostly co-expressed with β3 subunit Table 1 Antibodies used in the study Antibody Type Dilution Host Catalog number Producer Na/K-ATPase β1 Monoclonal 1:100 Mouse MA3-930 Thermo Fisher, Waltham, USA Na/K-ATPase β2 Polyclonal 1:50 Rabbit ANP-012 Alomone Labs, Jerusalem, Israel Na/K-ATPase β3 Polyclonal 1:200 Rabbit Abx111158 Abbexa, Cambridge, UK Na/K-ATPase α1′ Monoclonal 1:50 Mouse SC 21712 Santa Cruz Biotechnology, Dallas, USA Na/K-ATPase α1′′ Monoclonal 1:50 Mouse NB300-146 Novus, Littleton, USA Na/K-ATPase α2 Polyclonal 1:100 Rabbit OAAB03232 Aviva Systems Biology, San Diego, USA Na/K-ATPase α3 Monoclonal 1:50 Mouse NB300-540 Novus, Littleton, USA Carbonic anhydrase II Polyclonal 1:50 Rabbit HPA001550 Atlas Antibodies, Stockholm, Sweden NKCC2 Polyclonal 1:200 Rabbit OABB01331 Aviva Systems Biology, San Diego, USA Mineralocorticoid receptor′ Monoclonal 1:50 Mouse MA1-620 Thermo Fisher, Waltham, USA Mineralocorticoid receptor′′ Polyclonal 1:200 Rabbit PA5-81527 Thermo Fisher, Waltham, USA Vasopressin receptor type 2 Polyclonal 1:50 Rabbit Abx133128 Abbexa, Cambridge, UK ENaC Polyclonal 1:100 Rabbit PA1-920A Thermo Fisher, Waltham, USA Aquaporin-2 Polyclonal 1:50 Rabbit NB110-74682 Novus, Littleton, USA Aquaporin-4 Polyclonal 1:50 Rabbit AQP-004 Alomone Labs, Jerusalem, Israel Pendrin Polyclonal 1:50 Rabbit NBP1-60106 Novus, Littleton, USA NKCC1 Polyclonal 1:100 Rabbit AB59791 Abcam, Cambridge, UK NCC Polyclonal 1:100 Rabbit PA5-77816 Thermo Fisher, Waltham, USA the lateral plasma membrane (Fig. 4, insets).

Techniques: Expressing, Membrane, Fluorescence

Journal: Cell and tissue research

Article Title: "Reversed polarization" of Na/K-ATPase-a sign of inverted transport in the human endolymphatic sac: a super-resolution structured illumination microscopy (SR-SIM) study.

doi: 10.1007/s00441-019-03106-7

Figure Lengend Snippet: Fig. 7 SR-SIM (single optical section) of an epithelial villus in the intermediate portion of the human ES. Inset showing level of sectioning (*) of the ES. Epithelial cells co-express Na/K-ATPase α1 and β3 iso- forms in the apical cell membrane (a). A neighboring cell expresses only the β3 isoform. Inset shows framed area b at higher magnification. The nucleus contains α1 complexes (arrow), while the cytoplasm harbors both isoforms. The apical cell membrane framed (c) in a (b). Both α1 and β3 isoforms are expressed in the apical cytoplasm and coalesce in the cell membrane. Inset shows framed area at higher magnification

Article Snippet: At high magnification, Na/K-ATPase protein was seen to translocate from organelles near the cell nucleus to *α1 subunit mostly co-expressed with β3 subunit Table 1 Antibodies used in the study Antibody Type Dilution Host Catalog number Producer Na/K-ATPase β1 Monoclonal 1:100 Mouse MA3-930 Thermo Fisher, Waltham, USA Na/K-ATPase β2 Polyclonal 1:50 Rabbit ANP-012 Alomone Labs, Jerusalem, Israel Na/K-ATPase β3 Polyclonal 1:200 Rabbit Abx111158 Abbexa, Cambridge, UK Na/K-ATPase α1′ Monoclonal 1:50 Mouse SC 21712 Santa Cruz Biotechnology, Dallas, USA Na/K-ATPase α1′′ Monoclonal 1:50 Mouse NB300-146 Novus, Littleton, USA Na/K-ATPase α2 Polyclonal 1:100 Rabbit OAAB03232 Aviva Systems Biology, San Diego, USA Na/K-ATPase α3 Monoclonal 1:50 Mouse NB300-540 Novus, Littleton, USA Carbonic anhydrase II Polyclonal 1:50 Rabbit HPA001550 Atlas Antibodies, Stockholm, Sweden NKCC2 Polyclonal 1:200 Rabbit OABB01331 Aviva Systems Biology, San Diego, USA Mineralocorticoid receptor′ Monoclonal 1:50 Mouse MA1-620 Thermo Fisher, Waltham, USA Mineralocorticoid receptor′′ Polyclonal 1:200 Rabbit PA5-81527 Thermo Fisher, Waltham, USA Vasopressin receptor type 2 Polyclonal 1:50 Rabbit Abx133128 Abbexa, Cambridge, UK ENaC Polyclonal 1:100 Rabbit PA1-920A Thermo Fisher, Waltham, USA Aquaporin-2 Polyclonal 1:50 Rabbit NB110-74682 Novus, Littleton, USA Aquaporin-4 Polyclonal 1:50 Rabbit AQP-004 Alomone Labs, Jerusalem, Israel Pendrin Polyclonal 1:50 Rabbit NBP1-60106 Novus, Littleton, USA NKCC1 Polyclonal 1:100 Rabbit AB59791 Abcam, Cambridge, UK NCC Polyclonal 1:100 Rabbit PA5-77816 Thermo Fisher, Waltham, USA the lateral plasma membrane (Fig. 4, insets).

Techniques: Membrane

Journal: Cell and tissue research

Article Title: "Reversed polarization" of Na/K-ATPase-a sign of inverted transport in the human endolymphatic sac: a super-resolution structured illumination microscopy (SR-SIM) study.

doi: 10.1007/s00441-019-03106-7

Figure Lengend Snippet: Fig. 8 Confocal microscopy of the intermediate human ES showing expression of Na/K- ATPase β1 and NKCC2 (a). Many NKCC2-positive cells lack ATPase expression. Mineral cor- ticoid receptor expression in the ES (b). Both Na/K-ATPase-posi- tive and negative cells are posi- tive. SR-SIM showing heavy ac- tivity of Na/K-ATPase in the basolateral plasma membrane (filled arrow) and NKCC2 posi- tivity (c). Confocal microscopy show expression of NCC and ENaC (d, e)

Article Snippet: At high magnification, Na/K-ATPase protein was seen to translocate from organelles near the cell nucleus to *α1 subunit mostly co-expressed with β3 subunit Table 1 Antibodies used in the study Antibody Type Dilution Host Catalog number Producer Na/K-ATPase β1 Monoclonal 1:100 Mouse MA3-930 Thermo Fisher, Waltham, USA Na/K-ATPase β2 Polyclonal 1:50 Rabbit ANP-012 Alomone Labs, Jerusalem, Israel Na/K-ATPase β3 Polyclonal 1:200 Rabbit Abx111158 Abbexa, Cambridge, UK Na/K-ATPase α1′ Monoclonal 1:50 Mouse SC 21712 Santa Cruz Biotechnology, Dallas, USA Na/K-ATPase α1′′ Monoclonal 1:50 Mouse NB300-146 Novus, Littleton, USA Na/K-ATPase α2 Polyclonal 1:100 Rabbit OAAB03232 Aviva Systems Biology, San Diego, USA Na/K-ATPase α3 Monoclonal 1:50 Mouse NB300-540 Novus, Littleton, USA Carbonic anhydrase II Polyclonal 1:50 Rabbit HPA001550 Atlas Antibodies, Stockholm, Sweden NKCC2 Polyclonal 1:200 Rabbit OABB01331 Aviva Systems Biology, San Diego, USA Mineralocorticoid receptor′ Monoclonal 1:50 Mouse MA1-620 Thermo Fisher, Waltham, USA Mineralocorticoid receptor′′ Polyclonal 1:200 Rabbit PA5-81527 Thermo Fisher, Waltham, USA Vasopressin receptor type 2 Polyclonal 1:50 Rabbit Abx133128 Abbexa, Cambridge, UK ENaC Polyclonal 1:100 Rabbit PA1-920A Thermo Fisher, Waltham, USA Aquaporin-2 Polyclonal 1:50 Rabbit NB110-74682 Novus, Littleton, USA Aquaporin-4 Polyclonal 1:50 Rabbit AQP-004 Alomone Labs, Jerusalem, Israel Pendrin Polyclonal 1:50 Rabbit NBP1-60106 Novus, Littleton, USA NKCC1 Polyclonal 1:100 Rabbit AB59791 Abcam, Cambridge, UK NCC Polyclonal 1:100 Rabbit PA5-77816 Thermo Fisher, Waltham, USA the lateral plasma membrane (Fig. 4, insets).

Techniques: Confocal Microscopy, Expressing, Clinical Proteomics, Membrane

Journal: Cell and tissue research

Article Title: "Reversed polarization" of Na/K-ATPase-a sign of inverted transport in the human endolymphatic sac: a super-resolution structured illumination microscopy (SR-SIM) study.

doi: 10.1007/s00441-019-03106-7

Figure Lengend Snippet: Fig. 9 Proposed implication of a “reversed polarization” of Na/K-ATPase isoform expression in the epithelium of the human ES. Cylindrical cells (left) express αβ1 in the basolateral membrane, while the right cell ex- presses α1β3 in the apical plasma membrane. A high Na+ conductance in the apical plasma membrane is due to the presence of amiloride-sensitive Na+ channels (Kim et al. 2009). Na+ channels and Na/K-ATPase may be controlled by the hormone aldosterone (Furuta et al. 1999; Akiyama et al. 2010). It may also activate apical and basolateral K+ channels (Mori and Wu 1996; Wu and Mori 1996; Kim et al. 2015). A thiazide-sensitive Na/ Cl cotransporter was also detected in the apical membrane by Akiyama et al. (Akiyama et al. 2008)

Article Snippet: At high magnification, Na/K-ATPase protein was seen to translocate from organelles near the cell nucleus to *α1 subunit mostly co-expressed with β3 subunit Table 1 Antibodies used in the study Antibody Type Dilution Host Catalog number Producer Na/K-ATPase β1 Monoclonal 1:100 Mouse MA3-930 Thermo Fisher, Waltham, USA Na/K-ATPase β2 Polyclonal 1:50 Rabbit ANP-012 Alomone Labs, Jerusalem, Israel Na/K-ATPase β3 Polyclonal 1:200 Rabbit Abx111158 Abbexa, Cambridge, UK Na/K-ATPase α1′ Monoclonal 1:50 Mouse SC 21712 Santa Cruz Biotechnology, Dallas, USA Na/K-ATPase α1′′ Monoclonal 1:50 Mouse NB300-146 Novus, Littleton, USA Na/K-ATPase α2 Polyclonal 1:100 Rabbit OAAB03232 Aviva Systems Biology, San Diego, USA Na/K-ATPase α3 Monoclonal 1:50 Mouse NB300-540 Novus, Littleton, USA Carbonic anhydrase II Polyclonal 1:50 Rabbit HPA001550 Atlas Antibodies, Stockholm, Sweden NKCC2 Polyclonal 1:200 Rabbit OABB01331 Aviva Systems Biology, San Diego, USA Mineralocorticoid receptor′ Monoclonal 1:50 Mouse MA1-620 Thermo Fisher, Waltham, USA Mineralocorticoid receptor′′ Polyclonal 1:200 Rabbit PA5-81527 Thermo Fisher, Waltham, USA Vasopressin receptor type 2 Polyclonal 1:50 Rabbit Abx133128 Abbexa, Cambridge, UK ENaC Polyclonal 1:100 Rabbit PA1-920A Thermo Fisher, Waltham, USA Aquaporin-2 Polyclonal 1:50 Rabbit NB110-74682 Novus, Littleton, USA Aquaporin-4 Polyclonal 1:50 Rabbit AQP-004 Alomone Labs, Jerusalem, Israel Pendrin Polyclonal 1:50 Rabbit NBP1-60106 Novus, Littleton, USA NKCC1 Polyclonal 1:100 Rabbit AB59791 Abcam, Cambridge, UK NCC Polyclonal 1:100 Rabbit PA5-77816 Thermo Fisher, Waltham, USA the lateral plasma membrane (Fig. 4, insets).

Techniques: Expressing, Membrane, Clinical Proteomics