Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: Roles of LRRC26 as an auxiliary γ1-subunit of large-conductance Ca 2+ -activated K + channels in bronchial smooth muscle cells

doi: 10.1152/ajplung.00331.2019

Figure Lengend Snippet: Large-conductance Ca2+-activated K+ channel α-subunit (BKα) and BKγ1 form molecular complex on the cell surface in mouse bronchial smooth muscle cells (mBSMCs). A: coimmunoprecipitation assay was performed using rat BSMs (rBSMs). Lysates were precipitated with anti-BKα antibody and blotted using either anti-BKα or anti-BKγ1 antibody. Similar results were obtained from 3 independent experiments. Two rats were used for each experiment. Lane 1, lysates from rBSM; lane 2, samples precipitated with control resin that cannot bind to antibodies; lane 3, samples precipitated with resin that binds to anti-BKα antibody. B: total internal reflection fluorescence (TIRF) imaging of mBSMCs, in which BKα and BKγ1 were labeled with each antibody. Fluorescent signals from particles corresponding to BKα, BKγ1, and colocalization are shown in green, red, and yellow, respectively. Merged image was overlapped with a cell image. C: ratio of BKα particles localized alone or colocalized with BKγ1 to total BKα particles in mBSMCs (BKα alone, 134 particles and colocalized, 214 particles from 15 cells).

Article Snippet: Fluorescently labeled cells were observed by using confocal (AIR, Nikon, Tokyo, Japan) or total internal reflection fluorescence (TIRF) microscopes (Nikon).

Techniques: Co-Immunoprecipitation Assay, Fluorescence, Imaging, Labeling