Next we investigated TRPM8 channel activity under various menthol concentrations and determined the menthol dose response on the single channel level (Fig. 6D). We found that menthol at different concentrations affects TRPM8
activity by mainly altering the open probability of the channel (Fig. 6D). In the figure 6D values of Po observed at 100 mV were plotted against menthol concentrations, total 36 experiments were conducted and numbers of events analyzed
for each menthol concentration were in a range of 400–1500. We also studied the cold sensitivity of TRPM8 reconstituted into planar lipid bilayers. Figure 7 demonstrates representative current traces of TRPM8 in planar lipid bilayers activated
by lowering temperature from 23°C to 16°C. Total 12 independent experimentswere conducted. These experiments confirm that the TRPM8 protein reconstituted into artificial lipidic bilayer resembles the properties of the native channel
and can be successfully used for studies.