Figure 5.

Automated patch clamp analysis of ML252 interactions with Kv7 activators in Kv7.2/Kv7.3 heteromeric channels. (A and D) Drug perfusion paradigms to assess functional outcomes of interactions between ML252 and either ML213 or ICA-069673 (ICA-73). In (A), currents were first recorded in control (no drugs added), following a 5-min incubation in ML252 (at ∼IC50 as determined in Figure S4A), and finally after a combination of either activator (multiple concentrations) with ML252. Currents were measured at a pulse to +20 mV where channels are maximally activated, from a holding potential of −80 mV. In (D), currents were first recorded in control (no drugs added), following a 5-min incubation in ICA-73 (30 μm) or ML213 (3 μm), and finally after a combination of either activator with ML252 (multiple concentrations). (B and C) Concentration responses of Kv7.2/Kv7.3 channels to (B) ML213 or (C) ICA-73, administered alone or after preincubation in 1.5 μm ML252. (E and F) Concentration responses of Kv7.2/Kv7.3 to ML252 alone or after preincubation with (E) ML213 or (F) ICA-73. In panels B, C, E, and F, currents (measured at +20 mV) were normalized to response in control conditions in each respective wells containing up to 10 cells (n = 7–8 wells).