Nest predation is one of the most important mortality factors of birds. Field observations showed that tits (Paridae) produce hissing calls and, usually, have lower breeding losses than nesting Ficedula flycatchers, which do not make such calls. We hypothesise that differences in fledgling success can be directly attributed to the vocal reaction of tits. We tested experimentally whether the hissing calls can affect the behaviour of a potential predator, analysing the response of the Yellow-necked Mouse Apodemus flavicollis to playback of calls of three Parid species. The number of visits by mice to two types of cavities (with playback and control) was not significantly different, but the average time spent by mice in cavities with playback (3.9 s) was significantly shorter than in cavities without playback (26.3 s). This suggests that hissing behaviour of tits significantly changes the exploration activity of predators, which may ultimately increase the breeding success of this group of birds relative to the flycatchers.Significance statementNest predation is one of the most important mortality factors of small land birds, but some anti-predatory mechanisms are still poorly recognised. Numerous studies demonstrate that incubating tits make hissing sounds, when a predator is near, but despite almost a century of research, there is little evidence these calls indeed affect behaviour of predators. By using a simple laboratory experiment, we demonstrated that the hissing acoustic signals used by tits may change the behaviour of yellow-necked mice, which are an important predator of cavity-nesting birds in temperate forests. Intruding mice withdrew from cavities where hissing sounds were played back. Our results suggest that the hissing behaviour of tits can change the exploration activity of potential predators and may increase breeding success of this group of birds relative to the flycatchers, which stay silent when their nest is threatened.