Immunolocalization of H⁺/K⁺-ATPases in Capan-1 cells grown on permeable membranes.

<p><b>A</b>: The gastric H^<b>+</b>/K^<b>+</b>-ATPase α subunit (HKα1) was labeled with Calbiochem 119101 (polyclonal, against HKα1C-terminal) and Alexa 488 (green). <b>B</b>: The gastric H^<b>+</b>/K^<b>+</b>-ATPase β subunit (HKβ) was detected with Sigma A-274 (2G11, anti HKβ, monoclonal) and Alexa 488 (green). <b>C</b>: The non-gastric H^<b>+</b>/K^<b>+</b>-ATPase α subunit (HKα2) was stained with non-gastric HKα2 antibody (C384-M79) and Alexa 488 (green) Three pairs of images are shown. <b>D</b>: Example of a control image without primary antibodies but with F-actin marker (phalloidin Texas red). DAPI was used to stain the nucleus (blue). All bars are 10 μm, and images from at least 10 independent experiments are presented as both x-y and x-z scans. In x-y scan the left images are taken mid-way through the monolayer, the right images are taken close to the apical membrane. Dotted lines in x-y scans indicate where the x-z scan was taken.