May and Ben Parker took full custody of their nephew Peter after his parents were killed in a plane crash.

Later on, Ben was killed in a burglary, leaving May and Peter all by themselves, as she continued to raise him as her own son. Peter cared too much for her, so he forbade himself from revealing to her that he was the heroic Spider-Man, since he feared the shock of seeing her nephew as the very thing she despised (thanks to the Daily Bugle's smear campaign), might have been too much for her. Later on she became romantically involved with one of Spider-Man's most hated foes, Doctor Octopus, although their engagement fell through.

Recently, she found out Peter was indeed Spider-Man, when she visited him while he was recovering from a major battle with the villainous Morlun. She has also lost her home in Queens, after an attack from the Vibranium-skinned Charlie Weiderman. Iron Man allowed the Parkers to live in Stark Tower for a time during which May was courted by the Avengers' butler, Edwin Jarvis. During the Civil War Spider-Man revealed his secret identity to the world in support of the Superhuman Registration Act. Regretting his decision, Peter denounced the Act and fled with May and Mary Jane to hide out in a seedy motel. There, Aunt May was shot by an unknown sniper hired by the Kingpin (Wilson Fisk), taking advantage of knowing Spider-Man's identity. Hospitalized, her current condition was so critical that Peter searched every and all means to find a way to save her life. Choosing a deal with the demon Mephisto as his last resort, May is once again healthy and living in her Forest Hills home. However, Peter's marriage to Mary Jane has been undone in history as payment for May's restoration.