The PM density and stability of the HBH-CFTR-3HA variants in CFBE Tet-on cells on 24-well plates were measured by cell surface ELISA as reported previously [43 (link)]. Briefly, cell surface CFTR-3HA was labeled with anti-HA antibody (16B12, BioLegend, San Diego, CA, USA), biotinylated anti-mouse IgG (Funakoshi, Japan), and NeutrAvidin protein-HRP (Thermo Fisher, Waltham, MA, USA) and detected by the Amplex Red reagent (Thermo Fisher, Waltham, MA, USA). For the PM stability measurement, Cd or PYO was added to the cell culture medium during the 4 h’ chase at 37 °C incubation.
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