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Jane and Jim met in a puppetry course at the University of Maryland, College Park, where they were both undergraduates. At the time, Jane was a senior majoring in art education and Jim was a freshman studying to become a commercial artist. Nebel later became Henson's puppeteering partner on the television show Sam and Friends and the two eventually married.
Though Jane Henson was one of Jim Henson's first puppeteering partners, she seldom supplied voices. One of the few exceptions was in the Sesame Street song "Women Can Be". She performed considerably less after the birth of the couple's second child, Cheryl. However, she would occasionally serve as a background performer in various projects afterwards, with her last known performance occurring in episode 103 of The Muppet Show.
Jane sat on the Board of Directors of the Jim Henson Legacy. As president of The Jane Henson Foundation, she donated to Sesame Workshop, provided funding to the Jim Henson's Fantastic World exhibit and The Carter Center founded by U.S. President Jimmy Carter.