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The end of the Neon Genesis Evangelion left many questions as to the fate of NERV, the Children, the Angels, and many other plot factors unanswered, unexplained, or otherwise hanging. To answer a legion of now befuddled Evangelion fans, Gainax remade the ending into a two-part movie known as Death and Rebirth which was to supersede episodes 25 and 26 of the television series, which then led to this movie, Death and Rebirth itself being recreated into part one of The End of Evangelion.

In 2015 huge, tremendously powerful alien war machines return to Tokyo. The only hope for Mankind's survival lies in the Evangelion, a humanoid fighting machine developed by NERV, a special United Nations agency.

Lain Iwakura appears to be an ordinary girl, with almost no experience with computers or technology. Yet the sudden suicide of a schoolmate, and a number of strange occurrences, conspire to pull Lain into the world of the Wired, where she gradually learns that nothing is what it seems to be.

The protagonist Akira Fudo learns from his best friend, Ryo Asuka, that an ancient race of demons has returned to take back the world from humans. Ryo tells Akira that the only way to defeat the demons is to incorporate their supernatural powers, and suggests that he unite with a demon himself.

Third-year high school student Koyomi Araragi barely survives a vampire attack, thanks to the help of Meme Oshino, a strange man who lives in an abandoned building. Despite being able to save his humanity, several powers from vampirism remained in his body as side effects.