Figure 3
Demonstration of the ability of glafenine and selected derivatives to correct F508del-CFTR in primary human HBE cells. (A) F508del-CFTR functional expression in well-differentiated primary human bronchial epithelial (HBE) cells determined from the increase in short-circuit current. The basolateral membrane was permeabilized using nystatin, and an apical-to-basolateral chloride gradient was imposed. Representative Isc responses of primary HBE cells expressing F508del-CFTR to sequential addition of 10 µM forskolin, 50 µM genistein, and 10 µM CFTRinh-172 after 24 h preincubation with 0.1% dimethylsulfoxide (vehicle), glafenine and compounds 49, 53, 54, 55 and 56 (10 µM), or VX-809 (1 μM) (n = 4). As a control Trikafta is tested with using VX = 770 (100 nM) instead of genistein. The asterisks mark glafenine derivatives (49, 56) that give a response significantly above the control level. (B,C) This is represented in two graphs, first as the change in current and second as the percentage of VX-809 response. (D) Additionally, included are the traces for each compound attained from the Ussing chamber in this experiment. Data in panels A,B and C are presented as the means ± SEM, n = 4, *p < 0.05, **p < 0.01 and ***p < 0.001.
