paper n a bosc 23 retroviral packaging cells atcc cat  (ATCC)

Journal: The Journal of Biological Chemistry

Article Title: Pathogenic residue insertion in neuronal nicotinic receptor alters intra- and inter-subunit interactions that tune channel gating

doi: 10.1016/j.jbc.2024.107266

Figure Lengend Snippet: Expression and agonist-dependent single-channel activity of single and pairwise mutant receptors. A , radio-labeled epibatidine binding to intact Bosc 23 cells–expressing WT and mutant receptors was measured as described in . Results from two independent transfections, each in duplicate, are shown, with control binding defined as that measured for the WT receptor in the presence of epibatidine alone. Net cell surface binding is the difference between binding in the presence of epibatidine alone ( open bars ) and that in the presence of ACh ( gray bars ); binding in the presence of ACh corresponds to intracellular binding sites. B , single-channel recordings from the same of patch of cell membrane before and after addition of nicotine (final concentration 100 nM) to the bath solution for the indicated receptors. Traces are representative of three experiments per receptor. The electrical artifact during nicotine addition is removed. The cell attached patch configuration was maintained throughout with a membrane potential of −70 mV and display bandwidth of 2 kHz.

Article Snippet: Bosc 23 cells, an HEK293-derived cell line (CLS Cat# 300192/p777_HEK293, RRID:CVCL_0045) , were used to express WT and mutant receptors.

Techniques: Expressing, Activity Assay, Mutagenesis, Labeling, Binding Assay, Transfection, Control, Membrane, Concentration Assay