Supplementary Figure 3: Mouse forepaw does not move during cooling thermal stimulation under isoflurane anesthesia.

a, b, c, Three example, averaged Layer 2/3 cortical whole-cell recordings (black traces) from different mice during cold-thermal (32-22°C) stimulation of the forepaw (blue trace). the distance of paw movement (orange) was monitored with a movement sensor arm resting on top of the forepaw digits while the force was kept constant (green). The movement sensor was sensitive enough to record a slow (about 3 s duration) movement of the peltier element during cooling that moved the paw by about 1 μm. This slight movement did not cause responses in cortex as (i) cortical control recordings showed no response, data not shown; (ii) the same stimulator was used for single nerve afferent fibers recordings and did not evoked tactile responses in low threshold mechanoreceptors; (iii) the cold response starts before movement onset in the middle cell, see vertical red dashed line in (b). Horizontal marks next to V_m represent, (a) –56 mV, (b) –64 mV, (c) –64 mV.