ABCMdb

PMID: 11600438

Galietta LV, Jayaraman S, Verkman AS
Cell-based assay for high-throughput quantitative screening of CFTR chloride transport agonists.
Am J Physiol Cell Physiol. 2001 Nov;281(5):C1734-42., [PubMed]

Sentences

No. Mutations Sentence Comment

35 ABCC7 p.Gly551Asp Fischer rat thyroid (FRT) cells were transfected with wild-type or G551D CFTR and selected in 0.6 mg/ml zeocin. Login to comment 40 ABCC7 p.Gly551Asp In addition, a stably transfected Chinese hamster ovary (CHO) cell line was generated that expressed YFP-H148Q and was subsequently transfected with an episomal plasmid (pCEP4) containing the coding sequence for wild-type or G551D CFTR. Login to comment 53 ABCC7 p.Gly551Asp FRT cells stably expressing wild-type or G551D CFTR were cultured on Snapwell inserts (Costar) for 8-10 days to form epithelial monolayers with high electrical resistance (3-4 k⍀⅐cm2 ). Login to comment 58 ABCC7 p.Gly551Asp Cells stably expressing CFTR (wild type or G551D) and YFP-H148Q were plated in 96-well black microplates (Corning Costar) at a density of 20,000 cells/well using a Labsystems Multidrop apparatus for automated liquid delivery. Login to comment 86 ABCC7 p.Gly551Asp Activation of G551D CFTR, a mutant that is processed normally but is relatively Cl- impermeable at the plasma membrane (17), was studied in stably transfected FRT cells. Login to comment 108 ABCC7 p.Gly551Asp B: experiments as in A done in Fischer rat thyroid (FRT) cells expressing G551D CFTR. Login to comment 169 ABCC7 p.Gly551Asp We evaluated with the plate reader assay the efficacy of genistein, a known activator of CFTR (8, 21, 23), to induce halide transport in cells expressing wild-type and G551D CFTR. Login to comment 171 ABCC7 p.Gly551Asp Forskolin was added during the assay at the submaximal concentration of 250 nM for wild-type CFTR or the high concentration of 5 ␮M for G551D CFTR. Login to comment 176 ABCC7 p.Gly551Asp ABCC7 p.Gly551Asp In FRT cells stably expressing G551D CFTR, genistein alone did not activate halide transport, but subsequent addition of forskolin gave a small but significant activation of G551D CFTR at the higher genistein concentrations (Fig. 5B). Login to comment 195 ABCC7 p.Gly551Asp B: same as in A, except on FRT cells expressing G551D CFTR. Login to comment