Rudie, from Dandy's World, is transfem! (Headcanon)

Sharps, from Objectified, is transfem! (Canon)

Nevermind, it's only canon now.

Scag, from Regretevator, is transfem! (Canon)

Elliot, from Forsaken, is transfem! (Headcanon)

Madeline, from Celeste, is transfem! (Canon)

Fluttershy, from My Little Pony: Equestria Girls, is transfem! (Headcanon)

Shrimpo, from Dandy's World, is transfem! (Headcanon)

Split, from Regretevator, is transfem! (Headcanon)

Lest, from Arcane, is transfem! (Canon)

Marcille, from Delicious in Dungeon, is transfem! (Headcanon)

Dandicus "Dandy" Dancifer, from Dandy's World, is transfem! (Headcanon)

Hatred from Block Tales is transfem! (Headcanon)

Calypso from Block Tales is transfem! (Headcanon)

Akiyama Mizuki, from Project SEKAI, is transfem! (Implied/interpretation)

Fluttershy, from My Little Pony: Friendship is Magic, is transfem! (Headcanon)