2.5 Assessment of induction and emergence times

An isoflurane concentration of 1.4% was used to assess isoflurane anesthesia induction and emergence times. Mice were intraperitoneally injected with CNO (3 mg/kg) or vehicle 1 h before anesthesia induction. The isoflurane concentration in the anesthesia induction box was monitored and maintained at 1.4% during the process. The mouse was placed into the anesthetic chamber, and the cage was rotated to place the mouse on its back every 10 s. The LORR was defined when the mouse did not twist the body and place all four feet on the floor within 30 s. After a 30 min exposure to 1.4% isoflurane, the mouse was removed from the anesthetic chamber, and placed in a supine position in room air. Emergence time was defined as the duration from the time of removal from the isoflurane chamber to the time the mouse righted itself (all feet on the floor).