Effects of reversed sequential and random applications of various concentrations of DiC₈-PI(4,5)P₂ on homomeric Kv7.3^T channels. (A) Multichannel recording in an excised inside-out membrane patch from an HM1-CHO cell transfected with Kv7.3^T cDNA. The patch contained up to 11 channels with current levels. 1, 10, and 100 µM DiC₈-PI(4,5)P₂ were applied sequentially in ascending order and then in reverse (descending) order. Holding potential is 0 mV. (B) Multichannel recording of Kv7.3^T channel activity induced by applications of different concentrations of DiC₈-PI(4,5)P₂ in random order. Holding potential is 0 mV. (C) Mean (±SEM) single-channel open probabilities (P_open), corrected for channel numbers, from five such patches exposed to the same sequences of DiC₈-PI(4,5)P₂ applications as in A. (D) Stability plot analysis (bin width of 2 s) of the response of a patch of membrane to a 60-s application of 100 µM DiC₈-PI(4,5)P₂.