Whole-cell patch-clamp analysis of Krisanaklan Cl⁻ channel inhibition.

<p>A. CFTR currents measured at a holding potential of 0−80 mV and +80 mV (in steps of 20 mV) in FRT cells expressing CFTR. Currents were recorded without forskolin (basal), and with 10 µM forskolin without or with Krisanaklan (1∶2000 dilution). (right) Current/voltage (I/V) plot of mean currents at 400 ms. B. (left) CaCC currents recorded in control conditions, in the presence of Krisanaklan, and in the presence of Krisanaklan (1∶2000 dilution) and 100 µM CaCC_inh-A01. Currents were recorded at voltages of −100 to +100 mV in 20 mV steps from a holding potential of −50 mV. (right) I/V plot of mean currents at 500 ms. C. (left) Whole-cell TMEM16A Cl⁻ currents recorded at a holding potential of 0 mV and pulsing to voltages between −100 and +100 mV (in steps of 20 mV) in TMEM16A-expressing FRT cells. Measurements done in the absence and presence of Krisanaklan (1∶2000 dilution), and TMEM16A inhibitor T16A_inh-A01 (10 µM). (right) I/V plot of mean currents at 500 ms. D. Transepithelial transport of active components in Krisanaklan in T84 cells grown on a porous transwell insert. Percentage transport of Krisanaklan inhibitory compound(s) at 30 and 60 min measured by bioassay of Cl⁻ channel inhibition (mean ± S.E., n = 3).