Effect of cholesterol depletion for 15 min treatment with 10 mM M β CD and for 90 min treatment with 2.5 mM M βCD:cholesterol (10:1) on capsaicin-activated TRPV1. (A) Whole-cell patch-clamp recordings for TRPV1 treated with MβCD:cholesterol during application with 1 μM capsaicin from ramp experiments evaluated at a holding potential of −60mV, with extracellular NMDG (buffer 2), and intracellular sodium ions (buffer 3). Current densities were initially equal for both the control and treated cells, but the time-dependent switch of ion flow direction observed in control cells was inhibited by treatment with MβCD:cholesterol (n = 4, p < 0.05). (B) YO-PRO uptake was observed during stimulation of MβCD-treated cells with 1 μM capsaicin (extracellular buffer 8). The fluorescence uptake rate increased during activation with 1 μM capsaicin, and a significant difference in rate was observed, as the uptake rate after ∼20 s of stimulation was reduced by ∼50% in treated cells compared to control cells (n = 34–41, p < 0.001).