The present experiment was designed to determine if the absence of an auditory cue (i.e., a “tone-off” cue) would be an effective conditioned stimulus (CS) for delay eyeblink conditioning and to test if the medial geniculate nucleus (MGN) is part of the sensory pathway for tone-off conditioning. Rats were given paired or unpaired delay eyeblink conditioning to examine if responding to a tone-off CS was due to an associative process. An inactivation technique was performed on a separate group of rats to determine if the MGN is needed to express tone-off conditioning. The results showed that rats given paired conditioning acquired robust conditioned eyeblink responses (CRs) compared with rats given unpaired conditioning and that expression of tone-off elicited CRs was impaired when the MGN was inactivated. The findings suggest that tone-on and tone-off eyeblink conditioning may share a common neural pathway.