Generating CFBE Cell Lines for CFTR Mutant Studies

The human CF bronchial epithelial cell line CFBE41o- (CFBE), with a CFTR∆F508/∆F508 genotype, was obtained from D. Gruenert (University of California, San Francisco) and grown as reported previously [43 (link)]. CFBE Tet-on cells stably expressing HBH-WT-CFTR-3HA, HBH-R75Q-CFTR-3HA, or HBH-M470V-CFTR-3HA under a tetracycline-responsive promoter were generated by lentivirus transduction under puromycin (3 µg/mL) selection as reported previously [43 (link)]. Briefly, lentiviral particles expressing HBH-WT-, M470V-, or M470V-CFTR-3HA were produced in the HEK293T cells with the Lenti-X Packaging System (Takara Bio, Japan) following the manufacturer’s instructions. The cell lines were generated by transduction with lentiviral particles containing the inducible HBH-CFTR-3HA, followed by selection with puromycin for 2 weeks.
Cadmium nitrate tetrahydrate (Cd), pyocyanin (PYO), and doxycycline hydrochloride (Dox) were purchased from FUJIFILM Wako Chemicals (Japan). VX-809, VX-661, and VX-770 were purchased from AdooQ BioScience (Irvine, CA, USA). VX-445 and DMSO were purchased from Selleck Chemicals (Houston, TX, USA) and Sigma-Aldrich (St. Louis, MO, USA), respectively.