Journal: Scientific Reports

Article Title: Goblet Cell Hyperplasia Requires High Bicarbonate Transport To Support Mucin Release

doi: 10.1038/srep36016

Figure Lengend Snippet: Representative confocal microscope images show extent of expression and subcellular localization of TMEM16A, SLC26A4, carbonic anhydrase 2 (CA2), SLC12A2, and ATP12A (images taken from BE37 cells; similar results were obtained from BE63 cells). Whenever permitted by the combination of primary antibodies, acetylated tubulin and MUC5AC were also stained as markers of ciliated and goblet cells, respectively. Bronchial epithelia were kept under control conditions or treated with IL-4 for 72 hrs. Larger images: xy sections (scale bar: 20 μm). Inset: xz sections (scale bar: 10 μm). Images with a different scale of view are shown in .

Article Snippet: The following primary antibodies and dilutions were used: rabbit monoclonal anti-TMEM16A [SP31] (ab64085, Abcam) at 1:200, mouse IgG1 anti-CFTR (ab570, J.R. Riordan, University of North Carolina at Chapel Hill, and Cystic Fibrosis Foundation Therapeutics) at 1:250, mouse polyclonal anti-SLC26A4 (H00005172-A01, Abnova) at 1:200, rabbit polyclonal anti-SLC12A2 (HPA020130, Sigma-Aldrich) at 1:1000, rabbit monoclonal anti-CA2 [EPR5195] (ab124687, Abcam) at 1:500, mouse IgG1 anti-MUC5AC (NCL-HGM-45M1, Novocastra) at 1:100, mouse IgG2B anti-acetylated tubulin (7451, Sigma Aldrich) at 1:300, rabbit polyclonal anti-ATP12A (HPA039526, Sigma-Aldrich) at 1:400.

Techniques: Microscopy, Expressing, Staining

Journal: Scientific Reports

Article Title: Goblet Cell Hyperplasia Requires High Bicarbonate Transport To Support Mucin Release

doi: 10.1038/srep36016

Figure Lengend Snippet: For simplicity, the cartoon shows all channels and transporters within the same cell although some components (e.g. CFTR and TMEM16A) are localized in separate cell types. The NKCC1 transporter (SLC12A2) promotes the intracellular accumulation of Cl − that is then secreted through TMEM16A and CFTR Cl − channels. Bicarbonate is accumulated inside the cell by means of basolateral transporters and by conversion from CO 2 . Pendrin then mediates the exchange of extracellular Cl − with intracellular HCO 3 − . The apical membrane also contains the ATP12A K + /H + pump and possibly a K + channel. Secretion of K + could be the mechanism controlling the acidification of apical fluid by ATP12A.

Article Snippet: The following primary antibodies and dilutions were used: rabbit monoclonal anti-TMEM16A [SP31] (ab64085, Abcam) at 1:200, mouse IgG1 anti-CFTR (ab570, J.R. Riordan, University of North Carolina at Chapel Hill, and Cystic Fibrosis Foundation Therapeutics) at 1:250, mouse polyclonal anti-SLC26A4 (H00005172-A01, Abnova) at 1:200, rabbit polyclonal anti-SLC12A2 (HPA020130, Sigma-Aldrich) at 1:1000, rabbit monoclonal anti-CA2 [EPR5195] (ab124687, Abcam) at 1:500, mouse IgG1 anti-MUC5AC (NCL-HGM-45M1, Novocastra) at 1:100, mouse IgG2B anti-acetylated tubulin (7451, Sigma Aldrich) at 1:300, rabbit polyclonal anti-ATP12A (HPA039526, Sigma-Aldrich) at 1:400.

Techniques: