Dr. Markway, doing research to prove the existence of ghosts, investigates Hill House, a large, eerie mansion with a lurid history of violent death and insanity. With him are the skeptical young Luke, who stands to inherit the house, the mysterious and clairvoyant Theodora and the insecure Eleanor, whose psychic abilities make her feel somehow attuned to whatever spirits inhabit the old mansion. As time goes by it becomes obvious that they have gotten more than they bargained for as the ghostly presence in the house manifests itself in horrific and deadly ways.

Dr. John Markway, an anthropologist with an interest in psychic research, learns that Hill House, an old mansion in New England, has a reputation for evil and supposedly is filled with supernatural powers; and he decides to conduct an experiment there. Assisting him are two women he has carefully selected: Eleanor Vance, a lonely, withdrawn woman who supposedly had a supernatural experience at the age of 10 and has devoted her life to caring for her invalid mother; and Theodora, a bohemian of lesbian leanings and remarkable extrasensory perception. Luke Sannerson, a skeptic who stands to inherit the house, accompanies them. Almost immediately the quartet are subjected to thunderous poundings, hideous screeching, and other terrifying phenomena for which Markway can find no rational explanation. Eleanor feels that the house is calling to her; and she begins to treat it as a living object. At this point Dr. Markway's skeptical wife, Grace, arrives and, defying the ghost, tries to persuade her husband to give up his experiments. Eleanor, who has fallen in love with Markway, now loses all touch with reality; and the other members of the group decide that for her own safety she must leave. As she drives away she feels a force tugging at the steering wheel; then, suddenly, Grace appears in the road, and Eleanor, in attempting to avoid hitting her, swerves off the road and is killed by crashing into the same tree under which the first mistress of Hill House died.