Conversion of PI(4,5)P₂ to PI(4)P by a VSP inhibits TRPV1-WT with little or no inhibition of TRPV1-R721A in inside-out excised patches. (A) Representative time course measurements for currents in inside-out excised patches of cells transfected with TRPV1-WT or TRPV1-R721A alone (left) or in combination with VSP (right). The dotted lines represent zero current. The bars above the current indicate when capsaicin was added. For each experiment, 1 µM capsaicin was added to cells held at a potential of −60 mV during the time indicated by the red bar. Once current activation had reached steady-state, defined as time = 0 and denoted by the arrows, the voltage protocol shown in the figure inset was applied. The current at the end of each pulse at 100 mV (blue) and −60 mV (black) is shown. (B) Time course of current inhibition (mean ± SEM) upon initiation of the VSP activation protocol for TRPV1-WT (black, n = 5) and TRPV1-R721A (red, n = 10). (C) All-points plots representing the current remaining at −60 mV after 90 s of applying the VSP activation voltage protocol. The red points represent the data for each patch, and the black points with whiskers represent the mean ± SEM. The numbers in parentheses denote the number of patches for each condition. The asterisk indicates there is a significant difference between the two conditions (Mann–Whitney U test, two-tailed, P ≤ 0.01).