Throughout its range in Latin America the jaguar Panthera onca is threatened by habitat loss and fragmentation, and conflict with humans. Protected areas in Panama harbour some of the last remaining suitable habitat for jaguars and are vital to conservation. However, no previous studies had assessed which factors in particular affect the tolerance of rural Panamanians towards jaguars and National Park conservation, which is important to jaguar persistence. Whether these factors are consistent with previous research on human–carnivore coexistence is unclear. To address this we estimated the number of instances of depredation of cattle by jaguars, and assessed attitudes and perceptions of rural Panamanians. We conducted semi-structured interviews in two disparate study areas: Cerro Hoya National Park and Darién National Park. Depredation events were more frequent in the latter, but only residents of the former reported conflict between people and coyotes Canis latrans. Positive perceptions of jaguars and National Parks, and criticism of park management, increased with level of education and land ownership. Men were more open to receiving help on their farms to mitigate impacts of jaguars, and more tolerant of the presence of jaguars, than women. Residents from both study areas indicated high appreciation for their respective National Parks. We provide recommendations to improve community outreach and education initiatives, and suggest priority areas for future mitigation efforts concerning human–jaguar interactions in Panama.