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  • Homestar Runner:
    • Strong Bad started out as a villain who was always doomed to lose to Homestar. Now he's debatably the most popular character, and definitely one of the most recognisable.
    • To a lesser extent, there's Homsar. He first appeared as a one-shot gag to make fun of a typo, and over time, has been promoted up the ranks to become part of the main cast. HR's creators have joked about avoiding giving Homsar his own cartoon since they would never need to use him again.
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    • Same goes for Senor Cardgage.
    • Trogdor and Stinkoman (Strong Bad's blue-haired anime counterpart) are also very popular with the fans. They even appear together in the fifth episode of SBCG4AP!
    • Also Eh! Steve from the Show Within a Show Sweet Cuppin' Cakes. Strong Bad insists that the main character is Sherlock, some kind of "mix between a cow and a helicopter" who is constantly trying (and failing) to get a worm out of the ground, and that Eh! Steve is a background character who delivers his Catchphrase once an episode. Nevertheless, Eh! Steve is vastly more popular and a much more recognizable symbol of the show.
    • Teen Girl Squad is another good example.
    • Strong Bad Lampshades this, along with The Other Darrin in the email original. After the departure of "Original Bubs," his temporary replacement "Onion Bubs" became a fan favorite.
      • Onion Bubs! ONION BUUUUUBS!!!
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    • Played with in-universe in Strong Bad's commentary for an old King of Town toon. He hates every single thing about it except for the Blacksmith.
      Strong Bad: Oh, now this guy had some class. The Blacksmith, that guy was a good guy. I hung out with him after the cartoon; yeah, we had some good times...
  • Red vs. Blue: Considering how the series is already an Ensemble Cast, there's numerous examples.
  • From what you can tell by the amount of views for 6 of the Speed Paints for Fairy Foxes done by creator StickFreeks herself, Kai, Kiki, and Louie might be in that territory.
  • Cuddles may be the Series Mascot of Happy Tree Friends but Flippy and Flaky are undeniably the most popular characters. Unsurprisingly they're the Fan-Preferred Couple of the franchise.
  • Greg the Spider from Arby 'n' the Chief.
  • RWBY:
    • Velvet Scarlatina showed up in a single scene designed to show the audience the prejudice Faunus face on Remnant. She was such a huge hit with the fandom that she was fleshed out into a proper character, put on a team and given a recurring role in the show. Her combat outfit was even given an artists competition where fans could present their ideas, the winner being the design that was eventually used (albeit with a few modifications).
    • The chainsaw-wielding White Fang heavy who kicked Weiss' ass in "No Brakes" has been getting a surprisingly large amount of mileage.
  • If the Emperor Had a Text-to-Speech Device: Rogal Dorn, Fountain of Memes extraordinaire. His portrayal as The Comically Serious, entirely oblivious of the universe's insanity and utterly immune to others' snark (of which there is plenty) has made him beloved by the fanbase, to the point every single thing he says gets memetic status.
  • Missi from The Vampair is likable for her charming looks, her vindictive personality, and her singing voice.

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